Directory of Researchers

UNO Tomohide
Graduate School of Agricultural Science / Department of Agrobioscience
Professor
Biology
Last Updated :2024/01/31

Researcher Profile and Settings

Affiliation

  • <Faculty / Graduate School / Others>

    Graduate School of Agricultural Science / Department of Agrobioscience
  • <Related Faculty / Graduate School / Others>

    Faculty of Agriculture / Department of Agrobioscience

Teaching

Research Activities

Research Interests

  • P450
  • Insect

Research Areas

  • Environmental science/Agricultural science / Entomology
  • Life sciences / Applied biochemistry

Awards

  • Jul. 2014 日本学術振興会, ひらめき☆ときめきサイエンス推進賞, 科学研究費社会還元事業であるひらめきときめきサイエンスを継続的に行ってきたこと

    UNO TOMOHIDE

    Others

Published Papers

  • Shiori Takeji, Mai Okada, Shu Hayashi, Kengo Kanamaru, Yuichi Uno, Hiromasa Imaishi, Tomohide Uno

    Abstract CYP2C19 is a member of the human microsomal cytochrome P450 (CYP). Significant variation in CYP2C19 levels and activity can be attributed to polymorphisms in this gene. Wildtype CYP2C19 and 13 mutants (CYP2C19.1B, CYP2C19.5A, CYP2C19.5B, CYP2C19.6, CYP2C19.8, CYP2C19.9, CYP2C19.10, CYP2C19.11, CYP2C19.13, CYP2C19.16, CYP2C19.19, CYP2C19.23, CYP2C19.30, and CYP2C19.33) were coexpressed with NADPH‐cytochrome P450 reductase in Escherichia coli. Hydroxylase activity toward testosterone and progesterone was also examined. Ten CYP2C19 variants showed Soret peaks (450 nm) typical of P450 in the reduced CO‐difference spectra. CYP2C19.11 and CYP2C19.23 showed higher testosterone 11α, 16α‐/17‐ and progesterone 6β‐,21‐,16α‐/17α‐hydroxylase activities than CYP2C19.1B. CYP2C19.6, CYP2C19.16, CYP2C19.19, and CYP2C19.30 showed lower activity than CYP2C19.1B. CYP2C19.9, CYP2C19.10. CYP2C19.13, and CYP2C19.33 showed different hydroxylation activities than CYP2C19.1B. These results indicated that CYP2C19 variants have very different substrate specificities for testosterone and progesterone.

    Wiley, 10 Oct. 2023, Biopharmaceutics & Drug Disposition

    Scientific journal

  • Asuka MATSUI, Makoto TOKUSHIGE, Akira MIZOGUCHI, Kengo KANAMARU, Katsuhiko SAKAMOTO, Yuichi UNO, Tomohide UNO

    Biology Centre, AS CR, 29 Mar. 2023, European Journal of Entomology, 120, 93 - 104

    Scientific journal

  • Mako Sasao, Tomohide Uno, Risa Kitagawa, Asuka Matsui, Fumika Toryu, Akira Mizoguchi, Kengo Kanamaru, Katsuhiko Sakamoto, Yuichi Uno

    Springer Science and Business Media LLC, 21 Sep. 2022, Histochemistry and Cell Biology, 159 (2), 199 - 208

    Scientific journal

  • Tomohide Uno, Yusuke Ozakiya, Mako Sasao, Katsuhiko Sakamoto, Yasuo Yamauchi, Yuichi Uno, Kengo Kanamaru, Akira Mizoguchi

    Rab proteins are small GTP-binding proteins and are the largest family in the Ras GTPase superfamily and mediate vesicular transport in cells. Diverse insulin-like peptides, such as bombyxin, are synthesized in the brain and secreted into the haemolymph by the corpus allatum (CA). In the brain of Bombyx mori, Rabs are expressed in a specific area; however, which Rabs actually link the secretion of bombyxin remains unknown. A double-staining analysis of nine Rabs ( Rab1, 3, 6, 7, 14, 21, 26, 39 and X4) and bombyxin indicated that Rab3-, Rab7-, Rab39-and RabX4-immunohistochemical reactivity (ir) areas overlapped with bombyxin-ir in the brain and CA in B. mori, while Rab6-, Rab14-and Rab21-irs partially overlapped in the CA. Rab1-ir occurred in the other immunopositive areas in CA. Rab26-ir did not occur in the brain. Rab39-ir occurred in UNC104, Rab39- effector,-immunopositive neurons in the brain and CA. Thus, Rab3, 7, 39 and X4 may regulate the exocytosis of bombyxin.

    CZECH ACAD SCI, INST ENTOMOLOGY, 2021, EUROPEAN JOURNAL OF ENTOMOLOGY, 118, 307 - 314, English

    Scientific journal

  • Yoshihiro Shiomi, Makoto Yoshimura, Yuko Hori, Yuta Ohira, Kenji Nagahama, Tomoko Ozaki, Mineo Takei, Takao Tanaka, Tomohide Uno

    BACKGROUND: Anorexia is a serious problem in patients with gastric cancer who have undergone gastrectomy. Ghrelin, an orexigenic hormone primarily secreted from the stomach, has been proposed to prevent anorexia. Significant reduction in plasma ghrelin levels after gastrectomy may contribute to lack of appetite and weight loss. In this study, we investigated the effects of Z-505, a ghrelin receptor agonist, on anorexia after total gastrectomy (TG) in a rat model. METHODS AND MATERIALS: Male Sprague-Dawley rats were used to establish a TG model, and then sham-operated (control) and TG rats were randomly assigned to four subgroups receiving administration of Z-505 (100 mg/kg, p.o., once daily) or vehicle for 14 d from day 14 to day 27 after TG. The food intake, body weight, and fat weight were evaluated during the test period. Moreover, the neuronal activity in the hypothalamus was evaluated on day 21 to investigate the mechanism of action of Z-505. RESULTS: In TG rats, Z-505 significantly improved the decrease in cumulative food intake induced by the surgery over 14 d (TG + vehicle; 213.8 ± 15.3 g, n = 12 versus TG + Z-505; 258.2 ± 13.1 g, n = 14, P < 0.05). Z-505 also significantly increased fat weight and had a milder effect on body weight over 14 d. In addition, Z-505 significantly increased the number of c-Fos-positive cells in the hypothalamic arcuate nucleus (TG + vehicle; 17.8 ± 2.0, n = 12 versus TG + Z-505; 72.2 ± 11.8, n = 12, P < 0.001). CONCLUSIONS: Z-505 may be a useful therapeutic treatment for anorexia after TG.

    Feb. 2020, The Journal of surgical research, 246, 527 - 534, English, International magazine

    Scientific journal

  • Kohji Yamamoto, Takuya Tsubota, Tomohide Uno, Yutaro Tsujita, Shingo Yokota, Hideki Sezutsu, Kazuei Mita

    We had previously reported a prostaglandin E synthase (bmPGES) in the silkworm Bombyx mori that catalyzes the isomerization of PGH2 to PGE2. The present study aimed to provide a genome-editing characterization of bmPGES in B. mori. Results showed bmPGES gene disruption to result in a reduced content of PGE2. The change affected the expression of chorion genes and egg formation in silkworms. Collectively, the results indicated that bmPGES could be involved in reproduction of B. mori. Therefore, this study provides insights into the physiological role of bmPGES and PGE2 in silkworms.

    08 Jan. 2020, Biochemical and biophysical research communications, 521 (2), 347 - 352, English, International magazine

    Scientific journal

  • Functional characterization of insect-specific RabX6 of Bombyx mori

    Tomohide UNO, Yusuke OZAKIYA, Masayuki FURUTANI, Katsuhiko SAKAMOTO, Yuichi UNO, Kengo KANAMARU, Akira MIZOGUCHI

    Jan. 2019, Histochemistry and Cell Biology, 151, 187 - 198, English

    [Refereed]

    Scientific journal

  • Molecular and functional characterization of the American cockroach, Periplaneta americana, Rab5: the first exopterygotan low molecular weight ovarian GTPase during oogenesis

    Mohamed Elmogy, Amr.A Mohamed, Muhammad Tufail, Tomohide Uno, Makio Takeda

    The small Rab GTPases are key regulators of membrane vesicle trafficking. Ovaries of Periplaneta americana (Linnaeus) (Blattodea: Blattidae) have small molecular weight GTP/ATP-binding proteins during early and late vitellogenic periods of oogenesis. However, the identification and characterization of the detected proteins have not been yet reported. Herein, we cloned a cDNA en

    Oct. 2018, Insect science, 25 (5), 751 - 764, English

    [Refereed]

    Scientific journal

  • Metabolism of steroids by cytochrome P450 2C9 variants.

    Tomohide Uno, Ryosuke Nakano, Rie Kitagawa, Mai Okada, kengo Kanamaru, Shinji Takenaka, Yuichi Uno, Hiromasa Imaishi

    CYP2C9 is a human microsomal cytochrome P450c (CYP). Much variation in CYP2C9 levels and activity can be attributed to polymorphisms of this gene. Wild-type CYP2C9 and ten mutants were co-expressed with NADPH-cytochrome P450 reductase in Escherichia coli. The hydroxylase activities toward steroids were examined. CYP2C9.2, CYP2C9.3, CYP2C9.4, CYP2C9.16, CYP2C9.28, CYP2C9.48 and

    Aug. 2018, Biopharmaceutics & Drug Disposition, 39 (8), 371 - 377, English

    [Refereed]

    Scientific journal

  • Shun Tamaki, Mitsuhiko Yagi, Yuki Nishihata, Hideki Yamaji, Yasushi Shigeri, Tomohide Uno, Hiromasa Imaishi

    The aromatic compound p-hydroxybenzoate (PHBA) is an important material with multiple applications, including as a building block of liquid crystal polymers in chemical industries. The cytochrome P450 (CYP) enzymes are beneficial monooxygenases for the synthesis of chemicals, and CYP53A15 from fungus Cochliobolus lunatus is capable of executing the hydroxylation from benzoate to PHBA. Here, we constructed a system for the bioconversion of benzoate to PHBA in Escherichia coli cells coexpressing CYP53A15 and human NADPH-P450 oxidoreductase (CPR) genes as a redox partner. For suitable coexpression of CYP53A15 and CPR, we originally constructed five plasmids in which we replaced the N-terminal transmembrane region of CYP53A15 with a portion of the N-terminus of various mammalian P450s. PHBA productivity was the greatest when CYP53A15 expression was induced at 20°C in 2×YT medium in host E. coli strain ΔgcvR transformed with an N-terminal transmembrane region of rabbit CYP2C3. By optimizing each reaction condition (reaction temperature, substrate concentration, reaction time, and E. coli cell concentration), we achieved 90% wholecell conversion of benzoate. Our data demonstrate that the described novel E. coli bioconversion system is a more efficient tool for PHBA production from benzoate than the previously described yeast system.

    Korean Society for Microbiology and Biotechnology, 01 Mar. 2018, Journal of Microbiology and Biotechnology, 28 (3), 439 - 447, English

    [Refereed]

    Scientific journal

  • Metabolism of 7-ethoxycoumarin, flavanone and steroids by cytochrome P450 2C9 variants.

    Uno T, Nakano R, Kanamaru K, Takenaka S, Uno Y, Imaishi H

    Nov. 2017, Biopharm Drug Dispos, 38 (8), 486 - 493, English

    [Refereed]

    Scientific journal

  • Tomohide Uno, Masayuki Furutani, Katsuhiko Sakamoto, Yuichi Uno, Kengo Kanamaru, Akira Mizoguchi, Susumu Hiragaki, Makio Takeda

    Rab proteins are small monomeric GTPases/GTP-binding proteins, which form the largest branch of the Ras superfamily. The different Rab GTPases are localized to the cytosolic face of specific intracellular membranes, where they function as regulators of distinct steps in membrane trafficking. RabX4 is an insect-specific Rab protein that has no close homolog in vertebrates. There is little information about insect-specific Rab proteins. RabX4 was expressed in Escherichia coli and subsequently purified. Antibodies against Bombyx mori RabX4 were produced in rabbits for western immunoblotting and immunohistochemistry. Western blotting of neural tissues revealed a single band, at approximately 26kD. RabX4-like immunohistochemical reactivity was restricted to neurons of the pars intercerebralis and dorsolateral protocerebrum in the brain. Further immunohistochemical analysis revealed that RabX4 colocalized with Rab6 and bombyxin in the corpus allatum, a neuronal organ that secretes neuropeptides synthesized in the brain into the hemolymph. RabX4 expression in the frontal ganglion, part of the insect stomatogastric nervous system that is found in most insect orders, was restricted to two neurons on the outer region and did not colocalize with allatotropin or Rab6. Furthermore, RNA interference of RabX4 decreased bombyxin expression levels in the brain. These findings suggest that RabX4 is involved in the neurosecretion of a secretory organ in Bombyx mori.

    WILEY, Sep. 2017, ARCHIVES OF INSECT BIOCHEMISTRY AND PHYSIOLOGY, 96 (1), English

    [Refereed]

    Scientific journal

  • Tomohide Uno, Takeshi Yanase, Hiromasa Imaishi

    Aspergillus oryzae is a fungus widely used in traditional Japanese fermentation industries. Cytochrome P450 (CYP) proteins are ubiquitously distributed in nature and display a broad range of enzymatic activities. A novel CYP52 (CYP52G3) gene was found in A. oryzae. In this study, we report the functional characterization of CYP52G3. The recombinant protein was expressed heterologously in Escherichia coli, and its membrane fraction isolated. CYP52G3 showed activities for 7-ethoxycoumarin and alpha-naphtoflavone. Furthermore, CYP52G3 hydroxylated flavanone at the 4' and 6 position and metabolized some hydroxyl-flavanones and steroids. Bioconversion experiments indicated that CYP52G3 could convert flavanone and testosterone in a synthetic medium. The conversion rates of flavanone and testosterone at 24 H were 50% and 70%, respectively. These results support that CYP52G3 could prove a useful enzyme for the efficient production of new compounds from flavonoids and steroids. (C) 2016 International Union of Biochemistry and Molecular Biology, Inc.

    WILEY, May 2017, BIOTECHNOLOGY AND APPLIED BIOCHEMISTRY, 64 (3), 385 - 391, English

    [Refereed]

    Scientific journal

  • K. Yamamoto, Y. Ozakiya, T. Uno

    The aldo-keto reductase AKR2E4 reduces 3-dehydroecdysone to ecdysone in the silkworm Bombyx mori L. In this study, a quantitative polymerase chain reaction analysis revealed that the level of AKR2E4 mRNA was higher in the testes than in other tissues, and a western immunoblot analysis revealed that the AKR2E4 content in the testes was stage-specific from the fifth larval instar to the pupal stage. Immunohistochemical analysis showed that the AKR2E4 protein was present in cyst cells associated with sperm cells and spermatocytes. These results indicate that AKR2E4 plays an important role in 3-dehydroecdysone conversion to ecdysone and spermatogenesis in silkworm testes.

    Library of the University of Arizona, 2017, Journal of Insect Science, 17 (5), 1 - 3, English

    [Refereed]

    Scientific journal

  • Tomohide Uno, Masayuki Furutani, Chihiro Watanabe, Katsuhiko Sakamoto, Yuichi Uno, Kengo Kanamaru, Hiroshi Yamagata, Akira Mizoguchi, Makio Takeda

    In eukaryotic cells, Rab guanosine triphosphate-ases serve as key regulators of membrane-trafficking events, such as exocytosis and endocytosis. Rab3, Rab6, and Rab27 control the regulatory secretory pathway of neuropeptides and neurotransmitters. The cDNAs of Rab3, Rab6, and Rab27 from B. mori were inserted into a plasmid, transformed into Escherichia coli, and then subsequently purified. We then produced antibodies against Rab3, Rab6, and Rab27 of Bombyx mori in rabbits and rats for use in western immunoblotting and immunohistochemistry. Western immunoblotting of brain tissue revealed a single band at approximately 26 kDa. Immunohistochemistry results revealed that Rab3, Rab6, and Rab27 expression was restricted to neurons in the pars intercerebralis and dorsolateral protocerebrum of the brain. Rab3 and Rab6 co-localized with bombyxin, an insect neuropeptide. However, there was no Rab that co-localized with prothoracicotropic hormone. The corpus allatum secretes neuropeptides synthesized in the brain into the hemolymph. Results showed that Rab3 and Rab6 co-localized with bombyxin in the corpus allatum. These findings suggest that Rab3 and Rab6 are involved in neurosecretion in B. mori. This study is the first to report a possible relationship between Rab and neurosecretion in the insect corpus allatum.

    SPRINGER, Jul. 2016, HISTOCHEMISTRY AND CELL BIOLOGY, 146 (1), 59 - 69, English

    [Refereed]

    Scientific journal

  • Tomohide Uno, Chiho Izumi, Shinji Takenaka, Takeshi Yanase, Hiromasa Imaishi, Kengo Kanamaru, Hiroshi Yamagata, Yoshio Kaminishi, Takao Itakura

    We evaluated the metabolism of several herbicides and progesterone by two P450 proteins (CYP1A9 and CYP1C1) from Japanese eel (Anguilla japonica). Expression vectors harboring CYP1A9 and CYP1C1 sequences were introduced into Escherichia coli. E. coli membrane fractions were incubated with each substrate, and the metabolites were analyzed. CYP1A9 and CYP1C1 deethylated 7-ethoxycoumarin and phenacetin, and demethylated chlorotoluron, diuron, and linuron. CYP1C1 specifically hydroxlyated progesterone at the 6 beta and 16 alpha positions. Five amino acids of CYP1A9 related to substrate binding were selected for mutation analyses [CYP1A9(F128A), CYP1A9(F229A), CYP1A9(F263A), CYP1A9(V387A), and CYP1A9(I391A)]. Two variants, CYP1A9(F229A) and CYP1A9(F128A), changed the ratio of 16a hydroxyprogesterone to 613 hydroxyprogesterone. Among all the variants, CYP1A9(F263A) showed the highest activity towards substrates used. CYP1A9(V387A) and CYP1A9(I391A) showed higher activities than that of CYP1A9 toward progesterone. The substrate specificity of CYP1A9 may be altered by replacing an amino acid related to substrate binding. (C) 2015 Elsevier B.V. All rights reserved.

    ELSEVIER SCIENCE BV, Sep. 2015, ENVIRONMENTAL TOXICOLOGY AND PHARMACOLOGY, 40 (2), 360 - 368, English

    [Refereed]

    Scientific journal

  • Point mutation of cytochrome P450 2A6 (a polymorphic allele CYP2A6.25) confers new substrate specificity towards flavonoids

    UNO Tomohide, OGURA Chiaki, IZUMI Chihiro, NAKAMUJRA Masahiko, YANASE Takeshi, Hiroshi YAMAZAKI, ASHIDA Hitoshi, KANAMARU Kengo, YAMAGATA Hiroshi, IMAISHI Hiromasa

    Jul. 2015, Biopharmaceutics & Drug Disposition, 36 (8), 552 - 563, English

    [Refereed]

    Scientific journal

  • Tomohide Uno, Yuri Isoyama, Kazuki Sakamoto, Yuichi Uno, Katsuhiko Sakamoto, Kengo Kanamaru, Hiroshi Yamagata, Michihiro Takagi, Akira Mizoguchi, Makio Takeda

    Rab guanosine triphosphatases in eukaryotic cells are key regulators of membrane-trafficking events, such as exocytosis and endocytosis. Rab7 regulates traffic from early to late endosomes and from late endosomes to vacuoles/lysosomes. The Rab7-interacting lysosomal protein (RILP) was extracted from the silkworm, Bombyx mori (B. mori), and expressed in Escherichia coli (E. coli), followed by its purification. The glutathione sulfotransferase pull-down assay revealed that Rab7 of B. mori interacted with RILP of B. mori. We then produced antibodies against RILP of B. mori in rabbits for their use in Western immunoblotting and immunohistochemistry. Western immunoblotting of brain tissue for RILP revealed a single band, at approximately 50 kD. RILP-like immunohistochemical reactivity (RILP-ir) was restricted to neurons of the pars intercerebralis and dorsolateral protocerebrum. Furthermore, RILP-ir was colocalized with the eclosion hormone-ir and bombyxin-ir. However, RILP-ir was not colocalized with prothoracicotropic hormone-ir. These results were similar to those of Rab7 from our previous study. These findings suggest that RILP and Rab7 are involved in the neurosecretion in a restricted subtype of neurons in B. mori. Thus, our study is the first to report of a possible relationship between an insect Rab effector and neurosecretion.

    SPRINGER, Mar. 2014, HISTOCHEMISTRY AND CELL BIOLOGY, 141 (3), 311 - 320, English

    [Refereed]

    Scientific journal

  • Tomohide Uno, Yuri Isoyama, Kazuki Sakamoto, Yuichi Uno, Katsuhiko Sakamoto, Kengo Kanamaru, Hiroshi Yamagata, Michihiro Takagi, Akira Mizoguchi, Makio Takeda

    Rab guanosine triphosphatases in eukaryotic cells are key regulators of membrane-trafficking events, such as exocytosis and endocytosis. Rab7 regulates traffic from early to late endosomes and from late endosomes to vacuoles/lysosomes. The Rab7-interacting lysosomal protein (RILP) was extracted from the silkworm, Bombyx mori (B. mori), and expressed in Escherichia coli (E. coli), followed by its purification. The glutathione sulfotransferase pull-down assay revealed that Rab7 of B. mori interacted with RILP of B. mori. We then produced antibodies against RILP of B. mori in rabbits for their use in Western immunoblotting and immunohistochemistry. Western immunoblotting of brain tissue for RILP revealed a single band, at approximately 50 kD. RILP-like immunohistochemical reactivity (RILP-ir) was restricted to neurons of the pars intercerebralis and dorsolateral protocerebrum. Furthermore, RILP-ir was colocalized with the eclosion hormone-ir and bombyxin-ir. However, RILP-ir was not colocalized with prothoracicotropic hormone-ir. These results were similar to those of Rab7 from our previous study. These findings suggest that RILP and Rab7 are involved in the neurosecretion in a restricted subtype of neurons in B. mori. Thus, our study is the first to report of a possible relationship between an insect Rab effector and neurosecretion.

    SPRINGER, Mar. 2014, HISTOCHEMISTRY AND CELL BIOLOGY, 141 (3), 311 - 320, English

    [Refereed]

    Scientific journal

  • 宇野 知秀

    低分子量GTP(グアニンヌクレオチド三リン酸)結合タンパク質は,分子質量20-30kDa付近のGTP結合タンパク質の総称である。低分子量GTP結合タンパク質は,5つの主要なファミリー(Ras,Rho,Rab,ARF,Ran)に分類される(Wennerberg et al., 2005)。このうち,Rabファミリーは,最大のサブファミリーを形成し,細胞内の小胞の輸送に関与している。Rabの機能は,酵母の分泌の変異体を用いた初期の解析から明らかにされてきた(Novick et al., 1980; Salminen and Novick, 1987)。タンパク質は小胞体(endoplasmic reticulum; ER)で合成された後,ゴルジ体へ運ばれて,細胞内の小胞器官であるミトコンドリアやリソゾームへと運ばれる。また,ある種のタンパク質は細胞膜へと運ばれて,細胞外へと分泌される(exocytosis)。また,逆に細胞の外から中へ取り込まれる場合もある(endocytosis)。これらの輸送や分泌は,細胞内に存在する小胞により行われる。Rabは,これら小胞輸送の個々の段階に関与している。

    日本蚕糸学会, 2014, 蚕糸・昆虫バイオテック, 83 (1), 1_025 - 1_030, Japanese

  • Hamad Abu Zahra, Satoru Kuwamoto, Tomohide Uno, Kengo Kanamaru, Hiroshi Yamagata

    The cyclic nucleotides cGMP and cAMP have been reported to play key roles in the regulation of plant processes and responses. We have previously reported that several genes encoding flavonoid biosynthetic enzymes, including chalcone synthase (CHS) in soybean (Glycine max L), were induced by cGMP but not cAMP. The soybean genome contains nine CHS gene copies (GmCHS1-9). We investigated the responsiveness of several GmCHS genes to cGMP, CAMP, NO, and white light. Quantitative RT-PCR analysis showed that the transcript levels of GmCHS7 and GmCHS8 were increased by 3.6- and 3.8-fold, respectively, with cGMP whereas the transcript levels of GmCHS2 remained constant. Although CAMP had no effect on the transcript levels of the three genes, NO had an activation effect on all three. White light activated the three genes in a transient manner, with GmCHS2, GmCHS7, and GmCHS8 transcript levels increasing 3-fold after 3 h and decreasing to basal levels after 9 h. The GmCHS8 promoter contains several important cis-elements, including the G-box and H-box forming the Unit-I-like sequence and the MYB binding sequence, a target of the GmMYB176 transcription factor regulating the expression of GmCHS8. A transient gene expression assay revealed the activation of the Unit-I-like sequence, but not of the MYB binding sequence, by cGMP. The combination of G-box and H-box was necessary for cGMP responsiveness. Taken together, these results suggest that the Unit-I-like sequence in the promoters of GmCHS7 and GmCHS8 is a cGMP responsive cis-element in these genes and that NO exerts its effect via cis-elements other than the Unit-I-like sequence. (C) 2013 Elsevier Masson SAS. All rights reserved.

    ELSEVIER FRANCE-EDITIONS SCIENTIFIQUES MEDICALES ELSEVIER, Jan. 2014, PLANT PHYSIOLOGY AND BIOCHEMISTRY, 74 (1), 92 - 98, English

    [Refereed]

    Scientific journal

  • Metabolism of 7-ethoxycoumarin, safrole, flavanone and hydroxyflavanone by cytochrome P450 2A6 variants.

    UNO TOMOHIDE, 尾部 悠一郎, Ogura Chika, 山本 貢平, KANAMARU KENGO, YAMAGATA HIROSHI, IMAISHI HIROMASA

    多種の生体異物を代謝するヒトシトクロームP450 2A6の野生型および13種の変異型酵素をNADPH-シトクロームP450還元酵素とともに大腸菌で発現し、組換え酵素の7-エトキシクマリン、サロール、フラバノン、水酸化フラバノンの代謝を解析し、2A6ファミリーがフラボノイドを代謝することを明らかにした。

    Nov. 2013, Biopharm Drug Dispos., 34 (2), 87 - 97, English

    [Refereed]

    Scientific journal

  • Tomohide Uno, Kazuki Sakamoto, Yuri Isoyama, Susumu Hiragaki, Yuichi Uno, Kengo Kanamaru, Hiroshi Yamagata, Michihiro Takagi, Akira Mizoguchi, Makio Takeda

    Rab proteins are small GTPases that play essential roles in vesicle transport. In this study, we examined the expression of Rab proteins and neuropeptide hormones in the brain of the silkworm, Bombyx mori. We produced antibodies against B. mori Rab1 and Rab14 in rabbits. Immunoblotting of samples of brain tissue from B. mori revealed a single band for each antibody. Rab1 and Rab14 immunohistochemical labeling in the brain of B. mori was restricted to neurons of the pars intercerebralis and dorsolateral protocerebrum. Rab1, Rab7 and Rab14 co-localized with bombyxin. Rab1 and Rab7 co-localized with eclosion hormone. Rab1 co-localized with prothoracicotropic hormone. These results suggest that Rab1, Rab7 and Rab14 may be involved in neuropeptide transport in the brain of B. mori. This is the first report on the specificity of Rab proteins for the secretion of different neuropeptides in insects.

    SPRINGER, Feb. 2013, HISTOCHEMISTRY AND CELL BIOLOGY, 139 (2), 299 - 308, English

    [Refereed]

    Scientific journal

  • Tomohide Uno, Satoru Kaji, Tatsushi Goto, Hiromasa Imaishi, Masahiko Nakamura, Kengo Kanamaru, Hiroshi Yamagata, Yoshio Kaminishi, Takao Itakura

    Through the use of a number of bioconversion experiments we demonstrated that P450 proteins (CYP1A9 and CYP1C1) from Japanese eel (Anguilla japonica) metabolized a number of herbicides and the drug phenacetin. We performed bioconversion experiments in which substrates were added directly to incubation medium. The resulting metabolites were extracted and analyzed by high-performance liquid chromatography. Proteins CYP1A9 and CYP1C1 metabolized 50 nmol of the drug phenacetin to yield 12.1 and 1.1 nmol of product (acetaminophen), respectively. Further incubation of CYP1A9 with 50 nmol of the herbicides chlorotoluron, diuron, linuron, simazine, or atrazine yielded 16.5, 18.5, 7.3, 1.6, or 0.8 nmol of product, respectively. CYP1C1 also metabolized linuron, diuron, and simazine yield 5.4. 4.6, or 0.7 nmol of product, respectively. Next, polyclonal antibody was isolated by immunizing with two conjugated-peptides (amino acid residues 272-290 and 294-310) of CYP1A9. This antibody did not recognize human CYP1A2 or CYP1C1. Western blotting using the antibody revealed one band in the livers of Japanese eel and tilapia (Oreochromis niloticus). Theses results suggest that CYP1A9 and CYP1C1 metabolize herbicides, and that CYP1A9 is an useful biomarker of contamination when detected with this antibody. (C) 2011 Elsevier Inc. All rights reserved.

    ACADEMIC PRESS INC ELSEVIER SCIENCE, Oct. 2011, PESTICIDE BIOCHEMISTRY AND PHYSIOLOGY, 101 (2), 93 - 102, English

    [Refereed]

    Scientific journal

  • Tatsushi Goto, Hiroshi Moriuchi, Xuejun Fu, Tomoyo Ikegawa, Toshiyuki Matsubara, Gang Chang, Tomohide Uno, Kenichi Morigaki, Kunio Isshiki, Hiromasa Imaishi

    A number of studies have demonstrated that cytochrome P450 (P450) converts furanocoumarin derivatives into reactive molecules, which form covalent bonds to biomolecules. 5-Methoxypsoralen (5-MOP) is a natural furanocoumarin from apiaceous plants. In this study, we examined the effect on 5-MOP metabolism of single nucleotide polymorphisms (SNPs) in CYP2A13. We used Escherichia coli-generated recombinant enzymes of wild-type CYP2A13(star)1 and five variants, CYP2A13(star)4 (R101Q), CYP2A13(star)5 (F453Y), CYP2A13(star)6 (R494C), CYP2A13(star)8 (D158E), and CYP2A13(star)9 (V323L). In high-performance liquid chromatography analyses of 5-MOP metabolic products, CYP2A13(star)1 converted 5-MOP into 5-MOP dihydrodiol; K(m) and V(max) values of the reaction were 1.44 +/- 0.17 mu M and 4.23 +/- 0.36 nmol/(min . nmol P450), respectively. The generation of a dihydrodiol from 5-MOP implies that conversion by CYP2A13 causes toxicity due to the formation of covalent bonds with DNA or proteins. Most of the CYP2A13 variants could metabolize 5-MOP; K(m) values for CYP2A13(star)5, (star)6, (star)8, and (star)9 were 1.63 +/- 0.12, 1.36 +/- 0.10, 0.85 +/- 0.09, and 0.58 +/- 0.06 mu M, respectively, and V(max) values were 3.20 +/- 0.13, 4.69 +/- 0.13, 2.34 +/- 0.07, and 1.84 +/- 0.09 nmol/(min . nmol P450), respectively. However, the processing of 5-MOP by CYP2A13(star)4 was not detectable. Based on this data, we hypothesize that SNPs within the CYP2A13 gene affect metabolism of 5-MOP in humans.

    AMER SOC PHARMACOLOGY EXPERIMENTAL THERAPEUTICS, Dec. 2010, DRUG METABOLISM AND DISPOSITION, 38 (12), 2110 - 2116, English

    [Refereed]

    Scientific journal

  • Tomohide Uno, Keisuke Hata, Susumu Hiragaki, Yuri Isoyama, Le Thi Dieu Trang, Yuichi Uno, Kengo Kanamaru, Hiroshi Yamagata, Masahiko Nakamura, Michihiro Takagi, Makio Takeda

    Small GTPases of the Rab family are key regulators of membrane trafficking. We produced antibodies against the Rab7 protein of Bombyx mori (BRab7) in rabbits, and against the Rab11 protein of B. mori (BRab11) in mice. The antibodies recognized BRab7 and BRab11 proteins, but did not recognize other Rab proteins. Immunoblotting of samples from brain tissue of B. mori revealed a single band for each antibody. Rab11 was expressed in most tissues, whereas Rab7 was expressed in the brain, ovary, and testis. Immunohistochemical reactivity of Rab7 and Rab11 in the brain of B. mori was restricted to neurons of the pars intercerebralis and dorsolateral protocerebrum. Double-labeling experiments demonstrated that immunohistochemical reactivity of Rab7 co-localized with that of Rab11 and partially with that of Rab8. Immunohistochemical reactivity of Rab11 and Rab8 co-localized with that of PERIOD, one of the proteins associated with circadian rhythm. These findings suggest that Rab7, Rab8, and Rab11 are involved in protein transport in the neurons of the brain of B. mori and might play a role in the control of circadian rhythm.

    SPRINGER, Dec. 2010, HISTOCHEMISTRY AND CELL BIOLOGY, 134 (6), 615 - 622, English

    [Refereed]

    Scientific journal

  • Masataka Nakagawa, Megumi Ueyama, Hiroki Tsuruta, Tomohide Uno, Kengo Kanamaru, Bunzo Mikami, Hiroshi Yamagata

    Cucumisin is a subtilisin-like serine protease (subtilase) that is found in the juice of melon fruits (Cucumis melo L.). It is synthesized as a preproprotein consisting of a signal peptide, NH(2)-terminal propeptide, and 67-kDa protease domain. We investigated the role of this propeptide (88 residues) in the cucumisin precursor. Complementary DNAs encoding the propeptides of cucumisin, two other plant subtilases (Arabidopsis ARA12 and rice RSP1), and bacterial subtilisin E were expressed in Escherichia coli independently of their mature enzymes. The cucumisin propeptide strongly inhibited cucumisin in a competitive manner with a K(i) value of 6.2 +/- 0.55 nM. Interestingly, cucumisin was also strongly inhibited by ARA12 and RSP1 propeptides but not by the subtilisin E propeptide. In contrast, the propeptides of cucumisin, ARA12, and RSP1 did not inhibit subtilisin. Deletion analysis clearly showed that two hydrophobic regions, Asn(32)-Met(38) and Gly(97)-Leu(103), in the cucumisin propeptide were important for its inhibitory activity. Site-directed mutagenesis also confirmed the role of a Val(36)-centerd hydrophobic cluster within the Asn(32)-Met(38) region in cucumisin inhibition. Circular dichroism spectroscopy revealed that the cucumisin propeptide had a secondary structure without a cognate protease domain and that the thermal unfolding of the propeptide at 90 degrees C was only partial and reversible. A tripeptide, Ile(35)-Val(36)-Tyr(37), in the Asn(32)-Met(38) region was thought to contribute toward the formation of a proper secondary structure necessary for cucumisin inhibition. This is the first report on the function and structural information of the propeptide of a plant serine protease.

    AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC, Sep. 2010, JOURNAL OF BIOLOGICAL CHEMISTRY, 285 (39), 29797 - 29807, English

    [Refereed]

    Scientific journal

  • Tomohide Uno, Tsubasa Moriwaki, Yuri Isoyama, Yuichi Uno, Kengo Kanamaru, Hiroshi Yamagata, Masahiko Nakamura, Michihiro Takagi

    Rab GTPases are essential for vesicular transport, whereas adenosine triphosphate (ATP) is the most important and versatile of the activated carriers in the cell. But there are little reports to clarify the connection between ATP and Rab GTPases. A cDNA clone (Rab14) from Bombyx mori was expressed in Escherichia coli as a glutathione S-transferase fusion protein and purified. The protein bound to [H-3]-GDP and [S-35]-GTPgS. Binding of [S-35]-GTP gamma S was inhibited by guanosine diphosphate (GDP), guanosine triphosphate (GTP) and ATP. Rab14 showed GTP- and ATP-hydrolysis activity. The Km value of Rab14 for ATP was lower than that for GTP. Human Rab14 also showed an ATPase activity. Furthermore, bound [H-3]-GDP was exchanged efficiently with GTP and ATP. These results suggest that Rab14 is an ATPase as well as GTPase and gives Rab14 an exciting integrative function between cell metabolic status and membrane trafficking.

    ROYAL SOC, Jun. 2010, BIOLOGY LETTERS, 6 (3), 379 - 381, English

    [Refereed]

    Scientific journal

  • 食品の安全性評価へのバイオセンサーの利用

    今石 浩正, 後藤 達志, 宇野 知秀, 森垣 憲一

    2010, BRAIN TECHNO NEWS, 142, 21-25, Japanese

    [Refereed]

    Scientific journal

  • Makoto T. Fujiwara, Dongliang Li, Yusuke Kazama, Tomoko Abe, Tomohide Uno, Hiroshi Yamagata, Kengo Kanamaru, Ryuuichi D. Itoh

    Symmetric chloroplast division requires a prokaryote-derived division regulator protein MinD, whose subchloroplastic localization remains to be completely established. We investigated the localization and functionality of AtMinD1 (Arabidopsis thaliana MinD) fused with a dual hemagglutinin epitope (dHA) or a yellow fluorescent protein (YFP). AtMinD1-dHA, which successfully complemented the arc11/atminD1 mutant phenotype, was predominantly located at the envelope membrane and the mid-chloroplast constriction site. Meanwhile, AtMinD1-YFP was non-functional and showed suborganellar localization partly similar to that of AtMinD1-dHA. This prompts us to reevaluate earlier transgenic and transient expression studies using fluorescent protein-tagged AtMinD1.

    TAYLOR & FRANCIS LTD, Jul. 2009, BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 73 (7), 1693 - 1697, English

    [Refereed]

    Scientific journal

  • Susumu Hiragaki, Tomohide Uno, Makio Takeda

    Small GTPases of the Rab family act as essential regulators of vesicle transport pathways, including the exocytosis of neurohormones. These processes are not well-understood in insects. To address the physiological function of Rab proteins and their phosphorylation in insect neurosecretion, Rab8-like, prothoracicotropic hormone (PTTH)-like, and protein kinase C (PKC)-like immunohistochemical reactivities (-ir) were investigated in the brain of the American cockroach, Periplaneta americana. All the antibodies tested reacted with neurons in the pars intercerebralis, corpora cardiaca, and nervi corporis allati I. Double-labeling experiments demonstrated that all PTTH-ir were colocalized with Rab8-ir and PKC-ir in the pars intercerebralis, although exclusive reactivity was present to antisera against Rab8 or PKC. These findings support the notion that Rab8-like antigen is phosphorylated by PKC, and that this phosphorylation is involved in the axonal transport and secretion of PTTH in this species.

    SPRINGER, Mar. 2009, CELL AND TISSUE RESEARCH, 335 (3), 607 - 615, English

    [Refereed]

    Scientific journal

  • Uno T, Moriwaki T, Nakamura M, Matsubara M, Yamagata H, Kanamaru K, Takagi M

    Feb. 2009, Archives of insect biochemistry and physiology, 70 (2), 77 - 89

    [Refereed]

  • Phosphorylation of small GTPase Rab proteins from Bombyx mori (Lepidoptera: Bombycidae)

    Tomohide Uno, Keisuke Hata, Le Thi Dieu Trang, Susumu Hiragaki, Takuya Nakada, Masahiko Nakamura, Yuichi Uno, Hiroshi Yamagata, Kengo Kanamaru, Makio Takeda, Mamoru Matsubara

    Small GTPases of the Rab family act as essential regulators of vesicle transport pathways. Five Rab cDNA clones (BRab1, 7, 8, 11 and 14) from Bombyx mori were expressed in Escherichia coli as a thioredxin or glutathione sulfotransferase fusion protein. After purification, the fusion protein was tested for phosphorylation using protein kinase C (PKC). Results indicate that all of them were phosphorylated in vitro. The phosphorylation site of BRab1 was determined by mass-spectrometric analysis, which identified that Ser-17 of BRab1 was phosphorylated by PKC. Deletion and site-directed mutagenesis indicated that Ser-111 of BRab8, in addition to Ser-17, was newly phosphorylated. Further immunohistochemical analysis using antibodies against Rab8 indicated that there are some Rab8 immunoreactive cells close to the neuropeptide secreting cells. This result suggests that in insects Rab proteins are regulated by phosphorylation and at least some of them are involved in neuropeptide secretion.

    CZECH ACAD SCI, INST ENTOMOLOGY, 2009, EUROPEAN JOURNAL OF ENTOMOLOGY, 106 (4), 499 - 506, English

    [Refereed]

    Scientific journal

  • Ferulic Acid Production in the Brewing of Rice Wine (Sake)

    Tomohide Uno, Atsushi Itoh, Tetsuya Miyamoto, Masaharu Kubo, Kengo Kanamaru, Hiroshi Yamagata, Yukio Yasufuku, Hiromasa Imaishi

    The traditional Japanese alcoholic beverage sake is produced by fermentation of rice by Saccharomyces cerevisiae and Aspergillus oryzae. A. oryzae releases ferulic acid. an antioxidant, from steamed rice during the fermentation process. The concentration of ferulic acid increased with time during fermentation and the production rate peaked 9-12 days post inoculation. Analysis of the fermentation cultures of Aspergillus oryzae, by high-performance liquid chromatography (HPLC), revealed that p-coumaric acid induced an 18.9-fold increase in the level of ferulic acid. Furthermore, SDS-PAGE analysis revealed an increase or decrease in the level of specific proteins after the addition of p-coumaric acid to fermentation Cultures of Aspergillus oryzae. Ferulate esterase (FAE) activity was observed in the fermented sake ten days following the start of the fermentation process. These results suggest that the level of ferulic acid is regulated by the enzymes synthesized by A. oryzae during the sake brewing process.

    INST BREWING, 2009, JOURNAL OF THE INSTITUTE OF BREWING, 115 (2), 116 - 121, English

    [Refereed]

    Scientific journal

  • Tomohide Uno, Sota Okamoto, Satoko Masuda, Atsushi Itoh, Yuichi Uno, Masahiko Nakamura, Kengo Kanamaru, Hiroshi Yamagata, Hiromasa Imaishi

    P450 (cytochrome P450) enzymes catalyse the mono-oxygenation of a wide range of compounds such as steroids, fatty acids, vitamins and drugs. In the present paper we demonstrate a system for bioconverting diverse compounds [flavanone, DHEA (dehydroepiandrosterone) and 7-ethoxycoumarin] using P450 species expressed in Escherichia coli. First, we expressed four P450 species: rabbit CYP2B (MO family 2, subfamily B), fruitfly (Drosophila) CYP317A, rat CYP3A23 and mouse CYP2J5. Next, we added substrates directly to the incubation medium. The resulting metabolites were extracted and analysed by HPLC and spectrofluorimetry. The first substrate, 7-ethoxycoumarin, was de-ethylated by CYP2B; CYP2J5 and CYP3A23 showed weak activity, and CYP317A had no activity for 7-ethoxycoumarin. We next used flavanone, a flavonoid, as a substrate for these four MO species and other P450 species expressed previously. As a result, CYP2B, CYP2C43 and CYP2C29 catalysed flavanone 2-hydroxylation. CYP2A5 catalysed 2- and 4-hydroxylations. Finally, to produce diverse modified compounds, variants of CYP2A5 with point mutations were incubated with a steroid (DHEA) and an antioxidant (flavanone) in vivo. HPLC analysis indicated that two P450 species produced a 7-beta-hydroxy-DHEA and two P450 species produced a 2-alpha-hydroxy-DHEA. Four P450 species catalysed flavanone 2- and 4-hydroxylations. These results indicate that bioconversion by P450 is a useful technique to modify small molecules (steroids, coumarin and flavanone) and produce new, diverse hydroxylated compounds, which could be used for high-throughput screening for drug discovery.

    WILEY, Aug. 2008, BIOTECHNOLOGY AND APPLIED BIOCHEMISTRY, 50, 165 - 171, English

    [Refereed]

    Scientific journal

  • Tomohide Uno, Sota Okamoto, Satoko Masuda, Hiromasa Imaishi, Masahiko Nakamura, Kengo Kanamaru, Hiroshi Yamagata, Mohamed A. H. El-Kady, Yoshio Kaminishi, Takao Itakura

    We indicated that two P450s (1A9 and 1C1) from Japanese eel (Anguilla japonica) metabolized 7-ethoxycoumarin, 7-ethoxyresorufin, and flavanone. At first, we constructed expression vectors for two types of P450 (1A9 and 1C1). The reduced CO-difference spectra of Escherichia coli cells transformed with these plasmids showed Soret peaks (450 nm) that were typical of P450s. We performed bioconversion experiments in which substrates were added directly to incubation medium. The resulting metabolite(s) were extracted and analyzed by high-performance liquid chromatography and spectrofluorometer. Incubation of 50 nmol 7-ethoxyresorufin with P450 1C1 yielded 0.773 nmol of deethylated product, whereas 50 nmol 7-ethoxycoumarin resulted in 4.76 nmol. P450 1A9 metabolized 50 nmol of 7-ethoxyresorufin and 7-ethoxycoumarin to yield 6.54 and 20.9 nmol of deethylated product, respectively. Incubation of 50 nmol flavanone with P450 1C1 yielded 1.46 nmol and 0.69 nmol of products, whereas 50 mnol flavanone with P450 1A9 resulted in 1.10 nmol. In this system, 4'-hydroxy flavanones were formed by P450 1A9 and P450 1C1. P450 1A9 also metabolized 50 nmol of 17 beta-estradiol to yield 4.25 nmol of product. In this system, 2-hydroxy estradiol was formed by P450 1A9 using 17 beta-estradiol as a substrate. This study is the first to identify the substrates that P450 1C1 and 1A9 metabolize. (c) 2007 Elsevier Inc. All rights reserved.

    ELSEVIER SCIENCE INC, Apr. 2008, COMPARATIVE BIOCHEMISTRY AND PHYSIOLOGY C-TOXICOLOGY & PHARMACOLOGY, 147 (3), 278 - 285, English

    [Refereed]

    Scientific journal

  • Monoclonal antibody against rab8 from Bombyx mori (Lepidoptera : Bombycidae)

    Tomohide Uno, Takuya Nakada, Yuichi Uno, Kengo Kanamaru, Hiroshi Yamagata, Masahiko Nakamura, Michihiro Takagi

    Small GTPases of the Rab family are key regulators of membrane trafficking. Monoclonal antibodies are useful tools for identifying proteins that interact with other proteins and for examining their tissue distribution. We selected a monoclonal antibody against Rab8 of Bombyx Mori L. It specifically recognized amino acid residues 30-109, which are conserved among Rab8 proteins, and did not recognize any other Rab proteins. Western blotting using the antibody revealed one band in the brains of B. Mori and rat. Far-Western blotting analysis detected three proteins interacting with Rab8. These results indicate that this antibody is useful for clarifying the physiological function of Rab8 of B. Mori and other species. This is a report of a study on a monoclonal antibody against insect Rab protein.

    CZECH ACAD SCI, INST ENTOMOLOGY, Oct. 2007, EUROPEAN JOURNAL OF ENTOMOLOGY, 104 (4), 641 - 645, English

    [Refereed]

    Scientific journal

  • Tomohide Uno, Takuya Nakada, Sota Okamaoto, Masahiko Nakamura, Mamoru Matsubara, Hiromasa Imaishi, Hiroshi Yamagata, Kengo Kanamaru, Michihiro Takagi

    The Rob family of small GTPases are key regulators of membrane trafficking. Partially purified ROB from Bombyx mori (BRab8) was phosphorylated by protein kinase C in mammalian cells in vitro. To determine which of the seven serines and four threonines are phosphorylated, we generated deletion and site-directed mutants of BRab8, inserted them in Escherichia coli, partially purified the encoded fusion proteins by affinity chromatography, and examined their phosphorylation by protein kinase C in vitro. We found that Ser-132 of BRab8 was specifically phosphorylated by protein kinase C. In addition, Western blotting using an antiserum against BRab8 and in-gel staining for phosphorylated proteins revealed that BOB is phosphorylated in vivo.

    WILEY-BLACKWELL, Oct. 2007, ARCHIVES OF INSECT BIOCHEMISTRY AND PHYSIOLOGY, 66 (2), 89 - 97, English

    [Refereed]

    Scientific journal

  • Cloning and characterization of soybean GmGT-1 that binds to a light-responsive cis-element in ELIP promoter.

    Aoki, M, Uno, T, Kanamaru, K, Yamagata, H

    2007, Kobe Univ. Frontier Technology Forum 2007, 17, English

    [Refereed]

    Symposium

  • Development and characterization of photo-autophilic Arabidopsis cultured cells

    Li, T, Uno, T, Yamagata, H, Kanamaru, K

    2007, Kobe Univ. Frontier Technology Forum 2007, 18, English

    [Refereed]

    Symposium

  • Small GTP binding proteins; Rab GTPases from Bombyx mori.

    Moriwaki, T, Nakada, T, Uno, T, Kanamaru, K, Yamagata, H

    2007, Kobe Univ. Frontier Technology Forum 2007, 19

    [Refereed]

  • Bioconversion by functional P450 1A9 and 1C1of Anguillus japonica,

    UNO TOMOHIDE, Sota Okamoto, Satoko Masuda, Hiromasa Imaishi, Masahiko Nakamura, Kengo Kanamaru, Hiroshi Yamagata, Ei-Kady, Kaminishi, Takao Itakura

    2007, Comprehensive biochemistry and physiology C Toxicology Pharamacology, 147, 278 - 285, English

    [Refereed]

    Scientific journal

  • Tomohide Uno, Atsushi Nakao, Satoko Masuda, Yuuki Taniguchi, Kengo Kanamaru, Hiroshi Yamagata, Masahiko Nakamura, Hiromasa Imaishi, Kiyoharu Oono

    We developed a system for bioconverting diverse compounds using P450s produced in Escherichia coli. Vectors for the expressing various P450 cDNAs quickly and easily in E. coli were developed by using several restriction enzyme sites. Three types of P450 (2C2, 2C29, and 2D22) were produced using these plasmids. Substrates were directly added to the incubation medium and metabolized. To obtain pure product from the medium, we first tried production of P450 in synthetic medium. The amount of another P450 2C43 produced in the synthetic medium was similar to the amount produced in Luria broth (LB) medium. Next, estradiol, a steroid, was added as a substrate, incubated, and the metabolite was extracted and analyzed by high-performance liquid chromatography. The metabolite extracted from synthetic medium was purer than that obtained from LB medium. Three P450s (2C29, 2C2, and 2A4) metabolized testosterone at different positions. P450 2C29 metabolized 7-ethoxycournarin, androstendione, and dehydroepiandrosterone in this medium. P450s produced in the synthetic medium may be useful for producing various modified compounds for high-throughput screening.

    SPRINGER HEIDELBERG, Dec. 2006, JOURNAL OF INDUSTRIAL MICROBIOLOGY & BIOTECHNOLOGY, 33 (12), 1043 - 1050, English

    [Refereed]

    Scientific journal

  • Rice bifunctional alpha-amylase/subtilisin inhibitor: Cloning and characterization of the recombinant inhibitor expressed in Escherichia coli

    Teruyuki Yamasaki, Masaki Deguch, Toshiko Fujimoto, Takehiro Masumura, Tomohide Uno, Kengo Kanamaru, Hiroshi Yamagata

    The complete nucleotide sequences of the cDNA and its gene that encode a bifunctional alpha-amylase/subtilisin inhibitor of rice (Oryza sativa L.) (RASI) were analyzed. RASI cDNA (939 lip) encoded a 200-residue polypeptide with a molecular mass of 21,417 Da, including a signal peptide of 22 amino acids. Sequence comparison and phylogenetic analysis showed that RASI is closely related to alpha-amylase/subtilisin inhibitors from barley and wheat. RASI was found to be expressed only in seeds, suggesting that it has a seed-specific function. A coding region of RASI cDNA without the signal peptide was introduced into Escherichia coli and was expressed as a His-tagged protein. Recombinant RASI was purified to homogeneity in a single step by Ni-chelating affinity column chromatography and characterized to elucidate the target enzyme. The recombinant inhibitor had strong inhibitory activity toward subtilisin, with an equimolar relationship, comparable with that of native RASI, and weak inhibitory activity toward some microbial alpha-amylases, but not toward animal or insect alpha-amylases. These results suggest that RASI might function in the defense of the seed against microorganisms.

    TAYLOR & FRANCIS LTD, May 2006, BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 70 (5), 1200 - 1209, English

    [Refereed]

    Scientific journal

  • T Uno, A Nakao, Y Fujiwara, C Katsurauma, T Nakada, O Itoh

    Two partial cDNA clones (Protein kinase C alpha and Protein kinase C iota), each of which encoded a different member of PKC-protein family, were isolated using RT-PCR from mRNA of Bombyx mori. The full-length cDNAs were isolated using SMART-RACE. The cDNAs were expressed in HepG2 cells and the recombinant proteins were partially purified using an affinity chromatography. Protein kinase C alpha (BPKC alpha) showed a calcium-dependent kinase activity of histones. Whereas protein kinase C iota (BPKC iota) showed a calcium-independent activity. Bisindolyl maleimide 1, a PKC inhibitor, inhibited these kinase activities. Furthermore, in vitro BPKC alpha interacted and phosphorylated two proteins expressed in the brain of Bombyx mori. Rob protein, which plays important roles in the vesicle transport in the brain, and bMBD2/3, which is a methyl DNA-binding protein and regulates transcription. These results suggest that these PKCs phosphorylate various substrate proteins and function in the brain of Bombyx mori.

    WILEY-BLACKWELL, Feb. 2006, ARCHIVES OF INSECT BIOCHEMISTRY AND PHYSIOLOGY, 61 (2), 65 - 76, English

    [Refereed]

    Scientific journal

  • Biochemical characterization of Rab proteins from Bombyx mori,

    UNO TOMOHIDE, Tsubasa Moriwaki, Masahiko Nakamura, Mamoru Matsubara, Hiroshi Yamagata, Kengo Kanamaru, Michihiro Takagi

    The small GTPases known as Rab proteins are key regulators of membrane trafficking. We used RT-PCR to isolate cDNA clones of insect-specific Rab proteins (BRabN1 and BRabN2) showing low homology with known Rab proteins from other animals, from mRNA of Bombyx mori. These 2 Rabs were produced in Escherichia coli and purified. BRabN1 bound [(3)H]-GDP and [(35)S]-GTPgammaS with dis

    2006, Archives of Insect Biochemistry and Physiology, 70, 77 - 89, English

    [Refereed]

    Scientific journal

  • Expression, purification and characterization of methyl DNA binding protein from Bombyx mori.

    Uno T, Nomura Y, Nakamura M, Nakao A, Tajima S, Kanamaru K, Yamagata H, Iwanaga Y

    Aug. 2005, J Insect Sci, 5:8, 1-8

    [Refereed]

    Scientific journal

  • T Uno, A Nakasuji, M Shimoda, Y Aizono

    Suppression-subtractive hybridization was used to isolate cDNAs specifically expressed in the brain at the termination of pupal diapause in Agrius convolvuli. One of the isolated clones shows similarity to the cytochrome c oxidase subunit I (COX1) gene. The full-length cDNA was obtained from brain mRNA by rapid amplification of cDNA ends (RACE). The insert is 1.65 kb in length and has an open reading frame of 1.46 kb which encodes a putative protein of 486 amino acid residues. RT-PCR reveals that the mRNA increases dramatically at an early stage of diapause termination. Activity of cytochrome c oxidase in the brain also increases at the same time. The up-regulation of this gene suggests that expression of the COX1 gene and ATP synthesis are initiated in the brain in association with diapause termination. (C) 2003 Elsevier Ltd. All rights reserved.

    PERGAMON-ELSEVIER SCIENCE LTD, Jan. 2004, JOURNAL OF INSECT PHYSIOLOGY, 50 (1), 35 - 42, English

    Scientific journal

  • T Uno, S Hiragaki

    From a mRNA of the brain of Bombyx mori, we isolated 8 cDNA clones (BRabs), each of which encodes a different member of Rob-protein family. Four of them have more than 80% amino acid identity to the corresponding members of Drosophila Rob proteins. The other 4 proteins show low sequence similarity to any of the known Rob proteins. However, all of them contain the region conserved in rob protein. Using RACE (Rapid Amplification of cDNA ends), the one full-length cDNA clone (BRab14) was isolated. The clone was expressed in Escherichia coli as a glutathione S-transferase (GST) fusion protein. After purification, the fusion protein was cut with protease to remove GST-Tag and applied to a glutathione S-Sepharose column. The protein bound [H-3]-GDP with association constant of 1.02 x 10(11) M-1. Further, the protein was phosphorylated by protein kinase. This result suggests that Rob protein in the brain of Bombyx mori binds GDP or GTP and its function is regulated by phosphorylation.

    WILEY-LISS, Mar. 2003, ARCHIVES OF INSECT BIOCHEMISTRY AND PHYSIOLOGY, 52 (3), 130 - 138, English

    Scientific journal

  • Characteristics of psychrophilic alkaline phosphatase

    Y Ishida, H Tsuruta, ST Tsuneta, T Uno, K Watanabe, Y Aizono

    The phosphatase of a psychrophile (Shewanella sp.) was purified by ammonium sulfate fractionation, followed by sequential column chromatographies. The purified enzyme was electrophoretically homogeneous on native- and SDS-PAGE. Its molecular weight was 41,826 by its amino acid composition. The enzyme had its optimal pH for the activity at 9.8, and a broad substrate specificity to dephosphorylate ATP, pyrophosphate, glycerophosphate, and so on. Its activity was increased by metal ions including Mg2+, Mn2+, and Co2+. The maximal activity was observed at 40 degrees C, and the enzyme at 0 degrees C showed 39% of activity at 40 degrees C. The enzyme, however, tended to lose its activity at 20 degrees C and pH 9.8. These results indicated that purified enzyme was an alkaline phosphatase with characteristics; high catalytic efficiency at low temperature and gradual inactivation at an intermediate temperature.

    TAYLOR & FRANCIS LTD, Nov. 1998, BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 62 (11), 2246 - 2250, English

    [Refereed]

    Scientific journal

  • UNO Tomohide, UENO Mayumi, NAKAJIMA Ayumi, SHIRAI Yasuhito, AIZONO Yasuo

    From a brain cDNA library of Bombyx mori, we cloned cDNA for BRab, which encoded a 202-amino-acid polypeptide sharing 60-80% similarity with rab1 family members. To characterize its biochemical properties, cDNA for BRab was inserted into an expression vector (pGEX2T) and expressed in Escherichia coli as a glutathione S-trans-ferase (GST) fusion protein. The recombinant protein was purified to homogeneity with glutathione S-Sepharose. The purified GST-BRab bound [^<35>S]-GTPγS and [^3H]-GDP with association constants of 1.5×10^6M^<-1> and 0.58×10^6M^<-1>, respectively. The binding of [^<35>S]-GTPγS was inhibited with GTP and GDP, but with no other nucleotides. The GTP-hydrolysis activity was evaluated to be 5 m mole/min/mole of BRab. In the presence of 6 mM MgCl_2,bound [^<35>S]-GTPγS and [^3H]-GDP were exchanged with GTPγS most efficiently. These results suggest that BRab, having a higher affinity for GTP than GDP, converts from the GTP-bound state into the GDP-bound state by intrinsic GTP hydrolysis activity and returns to the GTP-bound state with the exchange of GDP with GTP.

    Japan Society for Bioscience, Biotechnology, and Agrochemistry, 23 Oct. 1998, Bioscience, biotechnology, and biochemistry, 62 (10), 1885 - 1891, English

  • Purification and some characteristics of phosphatase of a psychrophile

    H Tsuruta, ST Tsuneta, Y Ishida, K Watanabe, T Uno, Y Aizono

    The phosphatase of a psychrophile was purified by ammonium sulfate fractionation, and a sequence of chromatographies on DEAE-Cellulofine, butyl-Cellulofine, Sephacryl S-100, and Mono-Q columns, The purified enzyme preparation was found to be electrophoretically homogeneous on native-and SDS-PAGE, and its molecular mass was determined to be 38.4 kDa by MALDI-TOF mass spectrometry, Maximal activity was observed at 30 degrees C and pH 6.0, Furthermore, the activity of this enzyme at 0 and 5 degrees was 27 and 28%, respectively, of that at 30 degrees C, The enzyme was stable in the pH range of 6.0 to 8.0 and up to 20 degrees C. The enzyme was affected by metal ions; the activity was enhanced by Mg2+ and Ca2+ ions, but depressed by Zn2+ ions, Analysis of the amino acid composition indicated that this phosphatase contains no S-S bond, and only a few prolyl residues necessary to retain the rigid structure of a protein molecule, The phosphatase shows typical features of a cold enzyme; high catalytic activity at low temperature and rapid inactivation at an intermediate temperature.

    JAPANESE BIOCHEMICAL SOC, Feb. 1998, JOURNAL OF BIOCHEMISTRY, 123 (2), 219 - 225, English

    [Refereed]

    Scientific journal

  • Yoshio Imai, Tomohide Uno, Masahiko Nakamura

    cDNA for chimeric protein, P450(3P4), consisting of the amino-terminal 43 residues (the membrane-anchor region) of rabbit P450IIC14 and the remaining 447 residues of rabbit P450IIE1 was constructed, then cloned into expression vector pAAH5, and expressed in Saccharomyces cerevisiae AH22 cells under the control of yeast ADH1 promoter. P450(3P4) thus synthesized in the transformed yeast cells was partially purified, and its spectral and catalytic properties were examined. In the oxidized state P450(3P4) exhibited a high-spin type absorption spectrum even in the absence of a substrate. The reduced CO complex of the P450 showed a Soret absorption maximum at 452 nm. P450(3P4) catalyzed aniline p-hydroxylation, N-nitrosodimethylamine demethylation, benzphetamine N-demethylation, and laurate and caprate (ω-1)-hydroxylation in the reconstituted system containing the P450 and NADPH-P450 reductase. These results indicate that P450(3P4) preparation obtained from the transformed yeast cells has spectral and catalytic characteristics identical with those of P450IIE1 purified from rabbit liver microsomes, confirming the substrate specificity reported of P450IIE1. © 1990 Copyright, 1990 by the Journal of Biochemistry.

    Oxford University Press, 1990, Journal of Biochemistry, 108 (4), 522 - 524, English

    [Refereed]

    Scientific journal

MISC

  • ゲノム編集を利用して明らかにしたカイコ卵形成遺伝子の機能

    坪田拓也, 瀬筒秀樹, 山本幸治, 宇野知秀, 辻田裕太郎, 横田慎吾, 三田和英

    2019, 農研機構生物機能利用研究部門成果情報(Web), 2019

  • プロスタグランジンEのカイコ・コリオン遺伝子発現に与える影響

    広渡愛子, 坪田拓也, 宇野知秀, 瀬筒秀樹, 山本幸治

    2019, 日本蚕糸学会大会・蚕糸・昆虫機能学術講演会講演要旨集, 89th

  • 昆虫の低分子量GTP結合蛋白質(Rab)

    UNO TOMOHIDE

    2014, 蚕糸昆虫バイオテック, 83, 25 - 30, Japanese

    [Refereed]

    Introduction scientific journal

  • Tomohide Uno, Mayumi Ishizuka, Takao Itakura

    Cytochrome P450 (CYP) enzymes are members of the hemoprotein superfamily, and are involved in the mono-oxygenation reactions of a wide range of endogenous and exogenous compounds in mammals and plants. Characterization of CYP genes in fish has been carried out intensively over the last 20 years. In Japanese pufferfish (Takifugu rubripes), 54 genes encoding P450s have been identified. Across all species of fish, 137 genes encoding P450s have been identified. These genes are classified into 18 CYP families: namely, CYP1, CYP2, CYP3, CYP4, CYP5, CYP7, CYP8, CYP11, CYP17, CYP19, CYP20, CYP21, CYP24, CYP26, CYP27, CYP39, CYP46 and CYP51. We pinpointed eight CYP families: namely, CYP1, CYP2, CYP3, CYP4, CYP11, CYP17, CYP19 and CYP26 in this review because these CYP families are studied in detail. Studies of fish P450s have provided insights into the regulation of P450 genes by environmental stresses including water pollution. In this review, we present an overview of the CYP families in fish. (C) 2012 Elsevier B.V. All rights reserved.

    ELSEVIER SCIENCE BV, Jul. 2012, ENVIRONMENTAL TOXICOLOGY AND PHARMACOLOGY, 34 (1), 1 - 13, English

    [Refereed]

    Book review

  • Analysis of a knock-out mutant of an Arabidopsis T7 phage-type RNA polymerase, RpoTp (RpoT;3)

    Ryosaku Inagaki, Pyoyun Park, Michio Kanechi, Hirokazu Tsukaya, Kazuki Yanagida, Nozomu Sakurai, Hideyuki Suzuki, Daisuke Shibata, Tomohide Uno, Hiroshi Yamagata, Kengo Kanamaru

    OXFORD UNIV PRESS, 2007, PLANT AND CELL PHYSIOLOGY, 48, S84 - S84, English

    Summary international conference

  • Dynamism of chloroplast tRNA expression in Arabidopsis

    Mototsugu Kitamura, Kazuki Yanagida, Tomohide Uno, Hiroshi Yamagata, Kengo Kanamaru

    OXFORD UNIV PRESS, 2007, PLANT AND CELL PHYSIOLOGY, 48, S117 - S117, English

    Summary international conference

  • Functional involvement of cGMP and NO in light regulation of gene expression of soybean flavonoid-biosynthetic enzymes

    Kenji Suita, Maiko Mitsui, Tomohide Uno, Kengo Kanamaru, Hiroshi Yamagata

    OXFORD UNIV PRESS, 2007, PLANT AND CELL PHYSIOLOGY, 48, S65 - S65, English

    Summary international conference

  • Identification of regions functioning in substrate interaction of rabbit liver cytochrome P-450 (laurate (.OMEGA.-1)-hydroxylase).

    Uno Tomohide, Imai Yoshio

    The nucleotide sequence of cDNA for rabbit liver cytochrome P-450 (laurate (ω-1)-hydroxylase) was replaced with that for rabbit liver cytochrome P-450 (testosterone 16α-hydroxylase) in various regions coding for the amino acid sequence between residues 43 and 261. Six chimeric cDNAs thus constructed were cloned into expression vector pAAH5, and expressed in Saccharomyces cerevisiae AH22 cells under the control of yeast ADH1 promoter. Chimeric P-450s synthesized in the transformed yeast cells were purified partially and their catalytic and spectral properties were examined and compared with those of the chimeric P-450 which is considered to possess the same catalytic properties as the wild-type P-450. In the oxidized state the chimeric P-450s exhibited a low- and high-spin mixed-type absorption spectrum of cytochrome P-450 and the spectrum was converted to a typical high-spin type on addition of laurate or caprate, indicating the binding-of the fatty acids to the substrate site of the chimeric P-450s. However, the affinities of the fatty acids for the chimeras devoid of the sequence of P-450 (laurate (ω-1)-hydroxylase) in either of the regions spanning residues 90-125 and 210-261 were 10 to 20 times lower than those for the chimeras containing the sequence of the wild-type P-450 in both regions. The latter chimeras have about the same affinities as the chimera which is essentially the wild-type P-450. In the reconstituted system containing purified enzymes, the chimeras containing the sequence of the wild-type P-450 in both regions catalyzed (ω-1)-hydroxylation of the fatty acids, but their activities were 51-67% of that of the chimera which is essentially the wild-type P-450. The chimera which contains the sequence of the wild-type P-450 in the region covering residues 90-125 but not residues 210-261 was about one-tenth as active as the chimera which is essentially the wild-type P-450. On the other hand, the chimeras devoid of the sequence of the wild-type P-450 in the region spanning residues 90-125 were completely devoid of the hydroxylase activities. These results indicate that two segments of P-450 (laurate (ω-1)-hydroxylase) covering residues 90-125 and 210-262 constitute the substrate binding sites and cooperate to fix the fatty acid at an appropriate position on the P-450 molecule, and the former segment is essential to the hydroxylase activity. The reduced CO complex of the chimeric P-450s showed the typical absorption spectrum of cytochrome P-450. However, the CO complexes of the chimeras devoid of the sequence of the wild-type P-450 covering residues 90-125 were much more unstable to denaturation than those containing this sequence, suggesting that this sequence protects the conformation of the chimeric P-450s in the reduced state against denaturation.

    The Japanese Biochemical Society, 1989, J Biochem (Tokyo), 106 (4), 569 - 574, English

Books etc

  • BRAIN TECHNO NEWS 142

    今石 浩正, 後藤 達志, UNO TOMOHIDE, MORIGAKI KENICHI

    Others, 農研機構 生研センター, 2010, Japanese, 食品の安全性評価へのバイオセンサーの利用

    Scholarly book

Presentations

  • Transcriptional regulation in response to 5-aminolevulinic acid in Arabidopsis

    Imamura Nao, Tsiruyama Kaho, Iwamura Sakura, Sakamoto Minori, Kuroda Shuji, Uno Tomohide, KANAMARU KENGO

    JSPP, Mar. 2019, Japanese, Nagoya University, Domestic conference

    Oral presentation

  • Overexpression and characterization of Arabidopsis 5-aminolevulinic acid dehydratases in E. coli

    Kanbayashi Yuri, UNO TOMOHIDE, KANAMARU KENGO

    10th International Symposium of iBioK, Jan. 2019, English, Kobe University, International conference

    Poster presentation

  • 5-aminolevulinic acid responsive genes related to  increasing stress tolerance in Arabidopsis

    Tsuruyama Kaho, IMamura Nao, UNO TOMOHIDE, KANAMARU KENGO

    10th International Symposium of iBioK, Jan. 2019, English, Kobe University, International conference

    Poster presentation

  • 5-aminolevulinate up-regulated genes and their physiological effects in Arabidopsis

    KANAMARU KENGO, Tanaka Takahiko, Iwamura Sakura, Kuroda Shuji, Uno Tomohide

    第91回日本生化学会大会, Sep. 2018, Japanese, 京都国際会議場, Domestic conference

    Poster presentation

  • Function of 5-aminolevulinate membrane transporters in Arabidopsis

    Sakamoto Minori, Nakakita Takuma, Kuroda Shuji, Tanaka Takahiko, Uno Tomohide, KANAMARU KENGO

    第91回日本生化学会大会, Sep. 2018, Japanese, 京都国際会議場, Domestic conference

    Poster presentation

  • 昆虫の低分子量GTP結合蛋 白質Rabの機能解析

    UNO TOMOHIDE, 尾崎屋 悠祐, KANAMARU KENGO

    第62回日本応用動物昆虫学会大会, Mar. 2018, Japanese, 鹿児島大学郡元キャンパス, Domestic conference

    Oral presentation

  • 4-ヒドロキシ-5-アミノ-バレリン酸 (HAVA) によるシロイヌナズナの遺伝子発現応答

    IWAMURA SAKURA, TANAKA TAKAHIKO, SAKAMOTO MINORI, KURODA SHUJI, SAITO MASARU, IKEDA TOMOAKI, WATANABE SHIGEYUKI, UNO TOMOHIDE, KANAMARU KENGO

    日本農芸化学会2018年度大会, Mar. 2018, Japanese, Meijo University, Domestic conference

    Oral presentation

  • 植物における5-アミノレブリン酸関連トランスポーターの局在性と機能

    NAKAKITA TAKUMA, KONISHI MAHO, TERASHITA KAZUKI, YAMAGATA HIROSHI, UNO TOMOHIDE, KANAMARU KENGO

    ConBio2017, Dec. 2017, Japanese, 日本分子生物学会, 神戸ポートアイランド, Domestic conference

    Poster presentation

  • シロイヌナズナにおける5-アミノレブリン酸応答性遺伝子の機能的解析と分子機構

    TANAKA TAKAHIKO, DUAN CHEN, KURODA SHUJI, YAMAGATA HIROSHI, UNO TOMOHIDE, KANAMARU KENGO

    ConBio2017, Dec. 2017, Japanese, 日本分子生物学会, 神戸ポートアイランド, Domestic conference

    Poster presentation

  • 5-アミノレブリン酸近縁物質による 「負」の遺伝子発現制御

    IWAMURA SAKURA, TANAKA TAKAHIKO, SAKAMOTO MINORI, KURODA SHUJI, UNO TOMOHIDE, KANAMARU KENGO

    若手フロンティア研究会2017, Dec. 2017, Japanese, 神戸大学研究基盤センター, Domestic conference

    Poster presentation

  • 5-アミノレブリン酸近縁物質による 「正」の遺伝子発現制御

    TANAKA TAKAHIKO, DUAN CHEN, KURODA SHUJI, UNO TOMOHIDE, KANAMARU KENGO

    若手フロンティア研究会2017, Dec. 2017, Japanese, Kobe University, Domestic conference

    Poster presentation

  • 5-アミノレブリン酸(ALA)の膜輸送機構 〜変異株における細胞内ALA濃度と遺伝子発現への影響

    NAKAKITA TAKUMA, KURODA SHUJI, UNO TOMOHIDE, KANAMARU KENGO

    若手フロンティア研究会2017, Dec. 2017, Japanese, Kobe University, Domestic conference

    Poster presentation

  • 昆虫の脳に存在する蛋白質について

    UNO TOMOHIDE

    平成29年度西日本応用動物昆虫研究会・日本昆虫学会中国支部合同例会, Oct. 2017, Japanese, 神戸大学 瀧川記念学術交流会館大会議室, Domestic conference

    [Invited]

    Invited oral presentation

  • ALT1 and ALT2, 5-aminolevulinic acid (ALA) membrane transporters, in Arabidopsis

    NAKAKITA TAKUMA, KURODA SHUJI, YAMAGATA HIROSHI, UNO TOMOHIDE, KANAMARU KENGO

    8th International Symposium of iBioK, Feb. 2017, English, iBioK, 神戸大学, International conference

    Poster presentation

  • ALR1 and ALR2, 5-aminolevulinic acid (ALA)-responsive transcription factors, in Arabidopsis

    TANAKA TAKAHIKO, DUAN CHEN, KURODA SHUJI, YAMAGATA HIROSHI, UNO TOMOHIDE, KANAMARU KENGO

    8th International Symposium of iBioK, Feb. 2017, English, iBioK, 神戸大学, International conference

    Poster presentation

  • Molecular mechanisms of 5-aminolevulinic acid (ALA)-induced stress tolerance in Arabidopsis

    TANAKA TAKAHIKO, NAKAKITA TAKUMA, DUAN CHEN, KURODA SHUJI, YAMAGATA HIROSHI, UNO TOMOHIDE, KANAMARU KENGO

    第39回日本分子生物学会年会, Dec. 2016, Japanese, 日本分子生物学会, パシフィコ横浜, Domestic conference

    Poster presentation

  • Function of Arabidopsis 5-aminolevulinic acid transporter, ALT1, ALT2

    NAKAKITA TAKUMA, KONISHI MAHO, TERASHITA KAZUKI, YAMAGATA HIROSHI, UNO TOMOHIDE, KANAMARU KENGO

    第39回日本分子生物学会年会, Dec. 2016, Japanese, 日本分子生物学会, パシフィコ横浜, Domestic conference

    Poster presentation

  • 5-アミノレブリン酸誘導性転写応答による植物のマルチストレス耐性向上のメカニズム

    TANAKA TAKAHIKO, DUAN CHEN, KURODA SHUJI, YAMAGATA HIROSHI, UNO TOMOHIDE, KANAMARU KENGO

    日本農芸化学会関西支部第497回例会, Dec. 2016, Japanese, 日本農芸化学会関西支部, 神戸大学, Domestic conference

    Oral presentation

  • 5-アミノレブリン酸(ALA)の膜輸送機構 〜変異株における細胞内ALA濃度と遺伝子発現への影響

    NAKAKITA TAKUMA, KURODA SHUJI, KONISHI MAHO, TERASHITA KAZUKI, YAMAGATA HIROSHI, UNO TOMOHIDE, KANAMARU KENGO

    若手フロンティア研究会2016, Dec. 2016, Japanese, 神戸大学研究基盤センター, Domestic conference

    Poster presentation

  • 5-アミノレブリン酸(ALA)の分子機能 〜ALA誘導性遺伝子による植物のストレス耐性増強

    TANAKA TAKAHIKO, DUAN CHEN, KURODA SHUJI, YAMAGATA HIROSHI, UNO TOMOHIDE, KANAMARU KENGO

    若手フロンティア研究会2016, Dec. 2016, Japanese, 神戸大学研究基盤センター, Domestic conference

    Poster presentation

  • 昆虫の低分子量GTP結合蛋白質(Rab)について

    UNO TOMOHIDE

    第24回 WSフォーラム —タンパク質・ペプチド研究の現状と展望—, Nov. 2016, Japanese, 九州大学医系地区コラボステーションI 2階視聴覚ホール, Domestic conference

    [Invited]

    Invited oral presentation

  • 植物5-アミノレブリン酸(ALA)膜トランスポーターの局在性および機能解析

    NAKAKITA TAKUMA, KURODA SHUJI, KONISHI MAHO, TERASHITA KAZUKI, UNO TOMOHIDE, YAMAGATA HIROSHI, KANAMARU KENGO

    第11回トランスポーター研究会, Jul. 2016, Japanese, Kyoto University, 5-アミノレブリン酸(ALA)は、ヘムやクロロフィル等テトラピロール化合物生合成の共通前駆体である。植物では低濃度のALA投与で成長促進効果が、高濃度投与で光傷害誘導による枯死効果があることから、ALA含有肥料はすでに上市され,除草剤としても検討がされてきた。更に近年、中濃度のALA投与で様々な環境ストレスへの耐性が向上することがわかり、我々は新規植物活性化剤としての用途開発を視野にその分子機構について基盤研究を進めている。そのひとつがALAトランスポーターである。ALAトランスポーターの同定と機能解析は、植物アミノ酸輸送系の新たな一例となるだけでなく、ALAの生理効果を最大化して農業に活用する技術の開発基盤にもなる。 我々は動物ALAトランスポーターの情報や,推定される構造的・機能的特徴,局在性予測に基づき植物ALAトランスポーターの候補遺伝子, Domestic conference

    Poster presentation

  • Transcriptional Regulation and Improvement of Stress Tolerance Induced by 5-Amino Levulinic Acid

    KANAMARU KENGO, Duan Chen, Tanaka Takahiko, Nakakita Takuma, Masaru Saito, Fijimoto Takanori, Uno Tomohide, Yamagata Hiroshi

    第57回日本植物生理学会, Mar. 2016, Japanese, 岩手大学, Domestic conference

    Poster presentation

  • Possible 5-aminolevulinic acid (ALA) transporters (ALT) in Arabidopsis

    Nakakita Takuma, Kuroda Shuji, Konishi Maho, Miyake Michiru, Terashita Kazuki, Saito Masaru, Fujimoto Takanori, Uno Tomohide, Yamagata Hiroshi, KANAMARU KENGO

    The 7th International Symposium of Innovative BioProduction Kobe (iBioK), Jan. 2016, English, Kobe University, International conference

    Poster presentation

  • 5-aminolevulinic acid (ALA)-upregulated genes (ALU) and ALA-induced multi-stress tolerance in Arabidopsis

    Tanaka Takahiko, Duan Chen, Miyake Michiru, Saito Masaru, Fujimoto Takanori, Uno Tomohide, Yamagata Hiroshi, KANAMARU KENGO

    The 7th International Symposium of Innovative BioProduction Kobe (iBioK), Jan. 2016, English, Kobe University, International conference

    Poster presentation

  • 5-aminolevulinic acid (ALA)-responsive transcription factors (ALR) and regulatory gene expression in Arabidopsis

    Duan Chen, Kuroda Shuji, Nakajima Haruka, Nomura Hironari, Saito Masaru, Fujimoto Takanori, Uno Tomohide, Yamagata Hiroshi, KANAMARU KENGO

    The 7th International Symposium of Innovative BioProduction Kobe (iBioK), Jan. 2016, English, Kobe University, International conference

    Poster presentation

  • 植物をストレス耐性化する5-アミノレブリン酸の作用機構

    Duan Chen, Uno Tomohide, Yamagata Hiroshi, KANAMARU KENGO

    若手フロンティア2015, Dec. 2015, Japanese, Kobe University, Domestic conference

    Poster presentation

  • Molecular mechanism of 5-aminolevulinic acid (ALA)-dependent gene expression and increase of environmental stress tolerance in plant

    Duan Chen, Tanaka Takahiko, Nakakita Takuma, Konishi Maho, Masaru Saito, Fijimoto Takanori, Uno Tomohide, Yamagata Hiroshi, KANAMARU KENGO

    第38回日本分子生物学会年会, Dec. 2015, Japanese, 神戸ポートアイランド, Domestic conference

    Oral presentation

  • Molecular mechanism of 5-aminolevulinic acid (ALA)-dependent gene expression and increase of environmental stress tolerance in plant

    Duan Chen, Tanaka Takahiko, Nakakita Takuma, Konishi Maho, Masaru Saito, Fijimoto Takanori, Uno Tomohide, Yamagata Hiroshi, KANAMARU KENGO

    第38回日本分子生物学会年会, Dec. 2015, Japanese, 神戸ポートアイランド, 植物バイオマスの増産は,近年バイオリファイナリー産業の原料としての重要性が高まっており,植物の潜在的な生命力や環境適応力を最大限引き出す植物活性化物質の活用はその一法として有望である。 そこで我々は5-アミノレブリン酸(ALA)の生理活性に注目した。ALAはほぼ全ての生物が有するテトラピロール合成経路の中間基質で,合成されたヘムやクロロフィルは種々の酸化反応酵素や光合成集光色素の活性中心をなす。一方,植物種子や幼苗をALAで前処理しておくと塩や低温ストレスなど様々な環境ストレスへの耐性が長期間高まる現象が観察されている。この分子機構を解明すれば,ALAを肥料成分として以外に,塩害土壌などへの可耕地拡大,気象変動等による生育不良の改善など植物活性化物質として植物バイオマス増産に活用できる。 我々はまずトマトのストレス応答性転写因子からALA誘導性も, Domestic conference

    Poster presentation

  • 青色光/UV-AシグナルによるダイズELIP遺伝子の転写調節機構

    Akihiro Hohjo, Satoru Kuwamoto, UNO TOMOHIDE, KANAMARU KENGO, YAMAGATA HIROSHI

    日本農芸化学会2015年度大会, Mar. 2015, Japanese, 岡山大学, Domestic conference

    Oral presentation

  • ヌクレオシド三リン酸によるACC酸化酵素の活性化機構の解析

    Hiroki Iio, Takuma Maekawa, UNO TOMOHIDE, KANAMARU KENGO, YAMAGATA HIROSHI

    日本農芸化学会2015年度大会, Mar. 2015, Japanese, 岡山大学, Domestic conference

    Oral presentation

  • ククミシン遺伝子の果実特異的発現機構 -転写調節と形質転換トマトにおける発現解析-

    Okada Kaori, Miki Kohno, ATSUKO FURUKAWA, AIRI IEFUJI, UNO TOMOHIDE, KANAMARU KENGO, YAMAGATA HIROSHI

    日本農芸化学会2015年度大会, Mar. 2015, Japanese, Domestic conference

    Oral presentation

  • AtNAS1プロモーター中のNO応答性シスエレメントの解析

    Takuya Uchida, Tatsuya Ohara, Shunsuke Yano, Moeko Doi, UNO TOMOHIDE, KANAMARU KENGO, YAMAGATA HIROSHI

    日本農芸化学会2015年度大会, Mar. 2015, Japanese, 岡山大学, Domestic conference

    Oral presentation

  • 5-アミノレブリン酸(ALA)による植物遺伝子発現調節と環境ストレス耐性向上の分子機構

    Duan Chen, Tanaka Takahiko, Nakakita Takuma, Konishi Maho, Saitou Masaru, Fujimoto Hisanori, Uno Tomohide, Yamagata Hiroshi, KANAMARU KENGO

    BMB2015, Mar. 2015, Japanese, 神戸国際展示場, 植物バイオマスの増産は,近年バイオリファイナリー産業の原料としての重要性が高まっており,植物の潜在的な生命力や環境適応力を最大限引き出す植物活性化物質の活用はその一法として有望である。 そこで我々は5-アミノレブリン酸(ALA)の生理活性に注目した。ALAはほぼ全ての生物が有するテトラピロール合成経路の中間基質で,合成されたヘムやクロロフィルは種々の酸化反応酵素や光合成集光色素の活性中心をなす。一方,植物種子や幼苗をALAで前処理しておくと塩や低温ストレスなど様々な環境ストレスへの耐性が長期間高まる現象が観察されている。この分子機構を解明すれば,ALAを肥料成分として以外に,塩害土壌などへの可耕地拡大,気象変動等による生育不良の改善など植物活性化物質として植物バイオマス増産に活用できる。 我々はまずトマトのストレス応答性転写因子からALA誘導性も, Domestic conference

    Poster presentation

  • 青色光/UV-AによるダイズELIP遺伝子の転写調節機構

    Akihiro Hohjo, Satoru Kuwamoto, UNO TOMOHIDE, KANAMARU KENGO, YAMAGATA HIROSHI

    神戸大学研究基盤センター若手フロンティア研究会, Dec. 2014, Japanese, ダイズELIP(early light-inducible protein)遺伝子(GmELIP)の発現は青色光/UV-A照射により特異的に誘導される。GmELIPプロモーター中のGT-1-like boxとG boxの2つのシスエレメントが青色光/UV-A応答に必要かつ十分な制御配列である。転写因子GmGT-1がこれら両シスエレメントに結合することを既に報告したがG-box配列はbZIP型転写因子の標的配列でもある。そこで、今回は青色光/UV-AによるGmELIPの発現誘導におけるbZIP型転写因子の機能を解析した。 ダイズ光独立栄養培養細胞(SB-P細胞)中のGmELIPのUV-Aによる発現誘導はシクロヘキシミドによって強く阻害されたことから、この過程には新たなタンパク質合成が必要であることが示唆された。DNAマイクロアレイ分析による網羅的解析に, Domestic conference

    Poster presentation

  • ヌクレオシド三リン酸によるACC酸化酵素の活性化機構の解析

    HIROKI IIO, TAKUMA MAEKAWA, UNO TOMOHIDE, KANAMARU KENGO, YAMAGATA HIROSHI

    神戸大学研究基盤センター若手フロンティア研究会, Dec. 2014, Japanese, 神戸大学会館, エチレンは、花芽の形成や果実の追熟、器官脱離、老化、感染防御など様々な生理現象を制御している重要な植物ホルモンであり、メチオニンからS-アデノシルメチオニン、1-アミノシクロプロパン-1- カルボン酸(ACC)を経て合成される。ACC酸化酵素(ACO)はACCを酸化しエチレンに変換する最後の反応を触媒するが、その特性には不明な点が多い。本研究では、シロイヌナズナのAtACO2とトマトのLeACO1について新たな酵素学的特性を見出した。まず大腸菌で発現・精製したACOがアルカリ性で可逆的に失活することを明らかにした。次に、AtACO2の活性は12 mMヌクレオシド三リン酸(NTP)により約5倍に増加するのに対し、LeACO1は逆に約1/2に減少することを見出した。また、NTPのACCや各補因子に対するKm、Vmaxへの影響を調べた。一般的にNTPによる, Domestic conference

    Poster presentation

  • ククミシン遺伝子の果実特異的発現機構

    Kaori Okada, Miki Kohno, Atsuko Furukawa, Airi Iefuji, UNO TOMOHIDE, KANAMARU KENGO, YAMAGATA HIROSHI

    神戸大学研究基盤センター若手フロンティア研究会, Dec. 2014, Japanese, 神戸大学会館, メロン果実のセリンプロテアーゼ、ククミシンは果実の全可溶性タンパク質の10%以上を占める主要タンパク質で、受粉後10日前後の若い果実の果芯部でのみ発現する。ククミシンプロモーターの果実特異的発現能はトマトにおいても機能することを報告した。また、ククミシン遺伝子のプロモーター中に存在する、G-boxを含む20塩基の配列がククミシンの果実特異的な発現を制御するエンハンサーとして機能することが明らかにされ、その配列に結合する転写因子候補として、CmbZIP1とCmbZIP2が単離された。また、CmbZIP1とCmbZIP2自体が果実特異的に発現することも報告された。本研究では、CmbZIP1とCmbZIP2の果実内動態とDNA結合特性の解明を目的とした。受粉後10-15 日のメロン果実の核タンパク質中に上記のククミシンエンハンサーに結合するDNA結合タンパ, Domestic conference

    Poster presentation

  • AtNAS1プロモーター中のNO応答性シスエレメントの解析

    TAKUYA UCHIDA, TATSUYA OHARA, SHUNSUKE YANO, MOEKO DOI, UNO TOMOHIDE, KANAMARU KENGO, YAMAGATA HIROSHI

    神戸大学研究基盤センター若手フロンティア研究会, Dec. 2014, Japanese, 神戸大学会館, 一酸化窒素(NO)は植物において根の伸長や鉄の取り込み、気孔の閉鎖、植物ホルモンのシグナル伝達、病原菌感染や非生物的ストレスに対する適応応答等の様々な生理的過程に関与する重要なシグナル伝達分子である。動物に比べ、植物におけるNOシグナル伝達や遺伝子発現調節機構には不明な点が多い。本研究では、NOによる植物の遺伝子発現調節機構を転写レベルで明らかにすることを目的とした。シロイヌナズナの光独立栄養培養細胞(T87細胞)を用いたDNAマイクロアレイ解析により、NOによって発現が強く誘導される遺伝子としてニコチアナミン合成酵素遺伝子1(AtNAS1)が見出された。AtNAS1プロモーターの部分配列とGUSレポーター遺伝子の融合遺伝子をT87細胞のプロトプラストにPEG法で導入し、100 µM ニトロプルシドナトリウム(SNP、NO発生剤)で3時間処理後、GU, Domestic conference

    Poster presentation

  • Transcriptional control of fruit-secific expression of melon cucumisin

    Okada Kaori, Miki Kohno, Tomohide Uno, KANAMARU KENGO, YAMAGATA HIROSHI

    Plant Biology 2014, Jul. 2014, English, アメリカ植物生物学会, アメリカ・オレゴン州ポートランド・コンベンションセンター, Cucumisin is a subtilisin-like serine protease found in melon fruits. We reported the primary structure and characterization of cucumisin (1, 2). Cucumisin is synthesized in the central parts of young fruits and secreted into the juice; it comprises more than 15% of the total juice protein. A 20 bp enhancer element in the cucumisin promoter is responsible for the fruit-specific, International conference

    Poster presentation

  • シロイヌナズナRPOTmpの葉緑体光防護における役割

    KANAMARU KENGO, 山口 泰広, 李 棟梁, 一林 久雄, 野村 裕也, 宇野 知秀, 山形 裕士

    第55回日本植物生理学会年会, Mar. 2014, Japanese, 富山大学, Domestic conference

    Oral presentation

  • カイコ脳における低分子量GTP結合蛋白質(Rab)の免疫組織学的解析

    YAMAGATA HIROSHI, 古谷 昌之, 坂元 一樹, KANAMARU KENGO, TAKEDA MAKIO, 溝口 明, UNO TOMOHIDE

    平成26年度 蚕糸・昆虫機能利用学術講演会, Mar. 2014, Japanese, Domestic conference

    Oral presentation

  • 葉緑体・ミトコンドリア両局在性RNAPの光ストレス低減機能

    山口 泰広, 李 棟梁, 一林 久雄, 野村 裕也, 宇野 知秀, 山形 裕士, KANAMARU KENGO

    2013神戸大学研究基盤センター若手フロンティア研究会, Dec. 2013, Japanese, 神戸大学, Domestic conference

    Poster presentation

  • 青色光/UV-AによるダイズELIP遺伝子の発現制御機構

    Kuwamoto Satoru, Hamad Abu Zahra, Nogi Takasuke, Uno Tomohide, KANAMARU KENGO, YAMAGATA HIROSHI

    第36回日本分子生物学会年会, Dec. 2013, Japanese, 神戸国際会議場, Domestic conference

    Poster presentation

  • 青色光/UV-AによるダイズELIP遺伝子の発現制御機構

    桑本 知, Hamad Abu Zahra, 野木 貴祐, UNO TOMOHIDE, KANAMARU KENGO, YAMAGATA HIROSHI

    神戸大学研究基盤センター若手フロンティア研究会, Dec. 2013, Japanese, Domestic conference

    Poster presentation

  • メロン・ククミシン遺伝子プロモーターのトマトにおける発現解析

    Okada Kaori, Miki Kohno, Furukawa Atsuko, Hasegawa Sayako, Uno Tomohide, Kanamaru Kengo, YAMAGATA HIROSHI

    第36回日本分子生物学会年会, Dec. 2013, Japanese, 神戸国際会議場, Domestic conference

    Poster presentation

  • メロン・ククミシン遺伝子プロモーターのトマトにおける発現解析

    YAMAGATA HIROSHI, 岡田 香, 河野 美貴, 古川 温子, 長谷川 清子, UNO TOMOHIDE, KANAMARU KENGO

    神戸大学研究基盤センター若手フロンティア研究会, Dec. 2013, Japanese, Domestic conference

    Poster presentation

  • メロン・ククミシンの果実特異的発現を調節する転写因子CmbZIP1/2の特性解析

    Miki Kohno, Okada Kaori, Furukawa Atsuko, Uno Tomohide, Kanamaru Kengo, YAMAGATA HIROSHI

    第36回日本分子生物学会年会, Dec. 2013, Japanese, 神戸国際会議場, Domestic conference

    Poster presentation

  • メロン・ククミシンの果実特異的発現を調節する転写因子CmbZIP1/2の特性解析

    YAMAGATA HIROSHI, 河野 美貴, 岡田 香, 古川 温子, UNO TOMOHIDE, KANAMARU KENGO

    神戸大学研究基盤センター若手フロンティア研究会, Dec. 2013, Japanese, Domestic conference

    Poster presentation

  • シロイヌナズナRPOTmpの光ストレス応答における重要性

    山口 泰広, 李 棟梁, 宇野 知秀, 山形 裕士, KANAMARU KENGO

    第36回日本分子生物学会年会, Dec. 2013, Japanese, 神戸国際会議場, Domestic conference

    Poster presentation

  • Analysis of cis-element responsible for cGMP in the promoter of soybean chalcone synthase gene

    Hamad Abu Zahra, Satoru Kuwamoto, Tomohide Uno, Kengo Kanamaru, YAMAGATA HIROSHI

    第36回日本分子生物学会年会, Dec. 2013, English, 神戸国際会議場, Domestic conference

    Poster presentation

  • A cis-element responsible for cGMP in the promoter of the soybean chalcone synthase gene

    Hamad Abu Zahra, 桑本 知, UNO TOMOHIDE, KANAMARU KENGO, YAMAGATA HIROSHI

    神戸大学研究基盤センター若手フロンティア研究会, Dec. 2013, English, Domestic conference

    Poster presentation

  • 大腸菌で発現したウナギシトクロムP450(CYP1A9,CYP1C1)の機能解

    和泉 智保, 梶 悟, KANAMARU KENGO, YAMAGATA HIROSHI, 上西 由翁, 板倉 隆夫, UNO TOMOHIDE

    平成25年度日本水産学会秋季大会, Sep. 2013, Japanese, 三重大学, Domestic conference

    Oral presentation

  • Transcriptional control of fruit-specific expression of melon cucumisin.

    YAMAGATA HIROSHI, Miki Kono, Kaori Okada, Tomohide Uno, Kengo Kanamaru

    25th Congress of the Scandinavian Plant Physiology Society, Aug. 2013, English, Scandinavian Plant Physiology Society, Helsinger, Denmark, International conference

    Poster presentation

  • 免疫組織化学を用いたカイコ低分子量GTP結合タンパク質(Rab)の機能解析

    YAMAGATA HIROSHI, 坂元 一樹, 磯山 侑里, KANAMARU KENGO, 竹田 真生夫, 溝口 明, UNO TOMOHIDE

    日本蚕糸学会第83回大会, Mar. 2013, Japanese, つくば, Domestic conference

    Oral presentation

  • Involvement of Arabidopsis RPOTmp in chloroplast photoprotection

    KANAMARU KENGO, YAMAGUCHI YASUHIRO, Li D, ICHIBAYASHI HISAO, NOMURA HIRONARI, UNO TOMOHIDE, YAMAGATA HIROSHI

    第55回日本植物生理学会年会, Mar. 2013, Japanese, Toyama University, An Arabidopsis nuclear-encoded T7 phage-type RNA polymerase (NEP) named RPOTmp is a mitochondria/plastid dual targeting protein and fuction for the mitochondrial transcription of some respiratory chain complex genes, plastidic transcription of rrn operon during early development, and compensation of RPOTp function at some extent. We focused on the RPOTmp function for envirronme, Domestic conference

    Oral presentation

  • Function of Arabidopsis RPOTmp under light stress condition

    Yasuhiro Yamaguchi, Li Dongliang, Tomohide Uno, Hiroshi Yamagata, Hironari Nomura, KANAMARU KENGO

    第54回日本植物生理学会年会, Mar. 2013, Japanese, 岡山大学, Domestic conference

    Poster presentation

  • A study of the molecular function of 5-aminolevulinic acid for enhancement of salt-stress tolerance in higher plants

    Hironari Nomura, Haruka Nakajima, Jun Li, Shigeyuki Funada, Seiji Nishikawa, Tomohide Uno, Hiroshi Yamagata, KANAMARU KENGO

    第54回日本植物生理学会年会, Mar. 2013, Japanese, 岡山大学, Domestic conference

    Poster presentation

  • 5-アミノレブリン酸による植物塩ストレス耐性向上と遺伝子発現応答

    金丸 研吾, 中島 晴香, 山口 泰広, 李 潤, 舩田 茂行, 宇野 知秀, 山形 裕士, 野村 裕也

    第85回日本生化学会大会, Dec. 2012, Japanese, 福岡国際会議場, Domestic conference

    Poster presentation

  • 色素体局在性脂肪酸合成律速酵素の発現と機能

    小島 志織, 山本 俊佑, 能勢 琢也, 朴 杓允, 宇野 知秀, 山形 裕士, KANAMARU KENGO

    日本農芸化学会2011年度大会, Mar. 2011, Japanese, 京都女子大学, Domestic conference

    Oral presentation

  • クロルピリホス代謝に見られるヒトCYP2B6遺伝子多型の影響

    後藤 達志, 森 大気, 森垣 憲一, 宇野 知秀, 清水 泰博, 有澤 章, 今石 浩正

    日本農芸学会2011年度大会, Mar. 2011, Japanese, 日本農芸学会, 京都女子大, Domestic conference

    Poster presentation

  • 植物アセチルCoAカルボキシラーゼの発現と機能の解析

    小島 志織, 山本 俊佑, 能勢 琢也, 宇野 知秀, 山形 裕士, KANAMARU KENGO

    2010神戸大学研究基盤センター若手フロンティア研究会, Dec. 2010, Japanese, 神戸大学, Domestic conference

    Poster presentation

  • トマトにおける5-アミノレブリン酸の分子・生理機能

    中島 晴香, 小島 志織, 古閑 太郎, 都築 由起子, 舩田 茂行, 李 潤, 宇野 知秀, 山形 裕士, KANAMARU KENGO

    BMB2010(第33回日本分子生物学会年会 第83回日本生化学会大会), Dec. 2010, Japanese, 神戸ポートアイランド, Domestic conference

    Poster presentation

  • 色素体σ因子SIG6と発現相関する新規PPRの機能

    益 祐美子, 宇野 知秀, 山形 裕士, KANAMARU KENGO

    日本農芸化学会2010年度大会, Mar. 2010, Japanese, 東京大学, Domestic conference

    Oral presentation

  • 葉緑体発達に必須なシロイヌナズナPPRの分子機能解析.

    山本 俊佑, 小島 志織, 能勢 琢也, 稲垣 良作, 朴 杓允, 宇野 知秀, 山形 裕士, KANAMARU KENGO

    第32回日本分子生物学会年会, Dec. 2009, Japanese, パシフィコ横浜, Domestic conference

    Poster presentation

  • Molecular and physiological function of PPRTp, a pentatricopeptide repeat protein, in Arabidopsis (2)

    Shiori Kojima, Shunsuke Yamamoto, Takuya Nose, Tomohide Uno, Hiroshi Yamagata, KANAMARU KENGO

    WINPTech 2009, Dec. 2009, Japanese, 神戸大学, International conference

    Poster presentation

  • Molecular and physiological function of PPRTp, a pentatricopeptide repeart protein, in Arabidopsis (1)

    Shunsuke Yamamoto, Shiori Kojima, Takuya Nose, Pyoyun Park, Tomohide Uno, Hiroshi Yamagata, KANAMARU KENGO

    WINPTech2009, Dec. 2009, Japanese, 神戸大学, International conference

    Poster presentation

  • Molecular and physiological function of PPRS6a and PPRS6b, pentatricopeptide repeat proteins, in Arabidopsis

    Yumiko Masu, Tomohide Uno, Hiroshi Yamagata, KANAMARU KENGO

    WINPTech 2009, Dec. 2009, Japanese, 神戸大学, International conference

    Poster presentation

  • High-throughput Assay of Human P450 Activities by Using Immobilized Microsomes on Oxygen Sensor

    Gang Chang, 森垣 憲一, 達 吉郎, Uno Tomohide, 一色 邦夫, 今石 浩正

    薬物動態学会第24回年会, Nov. 2009, English, 国立京都国際会議場, Domestic conference

    Oral presentation

  • ヒトCYP2A13による食物由来成分の代謝における遺伝子多型の影響

    後藤 達志, 桧皮 貴史, 宇野 知秀, 一色 邦夫, 森垣 憲一, 今石 浩正

    第82回日本生化学会大会, Oct. 2009, Japanese, 神戸ポートアイランド, Domestic conference

    Oral presentation

  • シロイヌナズナ色素体accD遺伝子のRNA編集に関与するPPRの多面的効果

    小島 志織, 山本 俊佑, 能勢 琢也, 朴 杓允, 宇野 知秀, 山形 裕士, KANAMARU KENGO

    日本生化学会 2009年度大会, Oct. 2009, Japanese, 神戸国際会議場, Domestic conference

    Oral presentation

  • シトクロムP450遺伝子多型CYP2C9*2とCYP2C9*3が食品酸化に与える影響

    平井 洋輔, 後藤 達志, 宇野 知秀, 一色 邦夫, 森垣 憲一, 今石 浩正

    第82回日本生化学会大会, Oct. 2009, Japanese, 神戸ポートアイランド, Domestic conference

    Oral presentation

  • CYP1A1の遺伝子多型による食品成分の代謝活性変化

    桧皮 貴史, 後藤 達志, 宇野 知秀, 一色 邦夫, MORIGAKI KENICHI, 今石 浩正

    第82回日本生化学会大会, Oct. 2009, Japanese, 神戸ポートアイランド, Domestic conference

    Oral presentation

  • シロイヌナズナにおける5-ALAと関連分子の生理・分子作用

    川畑 絵里, 宇野 知秀, 山形 裕士, KANAMARU KENGO

    日本農芸化学会2009年度大会, Mar. 2009, Japanese, 福岡国際会議場, Domestic conference

    Poster presentation

  • Physiological and molecular functions of RPOTmp (RPOT;2) in Arabidopsis

    李 棟梁, 宇野 知秀, 山形 裕士, KANAMARU KENGO

    第50回日本植物生理学会年会, Mar. 2009, Japanese, 名古屋, Domestic conference

    Oral presentation

  • 色素体転写装置RPOTmp欠損変異株の生化学的解析

    李 棟梁, 宇野 知秀, 山形 裕士, KANAMARU KENGO

    2008神戸大学研究基盤センター若手フロンティア研究会, Dec. 2008, Japanese, 神戸大学, Domestic conference

    Poster presentation

  • 色素体転写・翻訳装置と機能相関する因子の探索と解析

    益 祐美子, 宇野 知秀, 山形 裕士, KANAMARU KENGO

    2008神戸大学研究基盤センター若手フロンティア研究会, Dec. 2008, Japanese, 神戸大学, Domestic conference

    Poster presentation

  • 色素体アセチルCoAカルボキシラーゼの発現と機能

    小島 志織, 山本 俊佑, 宇野 知秀, 山形 裕士, KANAMARU KENGO

    2008神戸大学研究基盤センター若手フロンティア研究会, Dec. 2008, Japanese, 神戸大学, Domestic conference

    Poster presentation

  • 色素体T7ファージ型RNAポリメラーゼと高い発現相関性をもつPPRTpの機能

    山本 俊佑, 能勢 琢也, 宇野 知秀, 山形 裕士, KANAMARU KENGO

    日本農芸化学会関西支部第457回講演会, Dec. 2008, Japanese, 日本盛 酒蔵通り煉瓦館, Domestic conference

    Oral presentation

  • 色素体T7ファージ型RNAポリメラーゼRpoTmpの機能解析

    李 棟梁, 一林 久雄, 宇野 知秀, 山形 裕士, KANAMARU KENGO

    第31回日本分子生物学会年会,第81回日本生化学会大会合同大会, Dec. 2008, Japanese, 神戸国際会議場, Domestic conference

    Oral presentation

  • Arabidopsis PPRTp is crucial for post-transcriptional regulation of plastidic accD gene.

    Shunsuke Yamamoto, Tomohide Uno, Hiroshi Yamagata, KANAMARU KENGO

    WINPTech 2008, Nov. 2008, Japanese, 神戸大学, International conference

    Poster presentation

  • シロイヌナズナ5-ALAデヒドラターゼの生理機能および 酵素学的性質の解析

    天尾 雅, 宇野 知秀, 山形 裕士, KANAMARU KENGO

    2007神戸大学研究基盤センター若手フロンティア研究会, Dec. 2007, Japanese, 神戸大学, Domestic conference

    Poster presentation

  • カイコGTP結合タンパク質(Rab)の機能解析

    森脇 翼, 中田 拓哉, 宇野 知秀, 金丸 研吾, 山形 裕士

    平成19年度蚕糸・昆虫利用学術講演会, Apr. 2007, Japanese, Domestic conference

    Oral presentation

  • 葉緑体tRNAは動的に発現調節されている

    北村 元嗣, 柳田 一樹, 宇野 知秀, 山形 裕士, 金丸 研吾

    日本植物生理学会2007年度年会, Mar. 2007, Japanese, 松山, Domestic conference

    Oral presentation

  • 高等生物のP450大腸菌発現系を用いたバイオコンバージョンシステムとその応用

    岡本 聡太, 宇野 知秀, 増田 智子, 伊東 鎮, 桝井 孝一, 山本 高明, 佐々木 彩子, 中西 昭二, 小野 雄介, 堀本 智仁, 金丸 研吾, 山形 裕士, 今石 浩正

    日本薬学会2007年度会, Mar. 2007, Japanese, 富山, Domestic conference

    Oral presentation

  • 光シグナル、cGMPおよびNOによるダイズフラボノイド合成系酵素遺伝子群の発現調節

    吹田 憲治, 三井 麻衣子, 宇野 知秀, 金丸 研吾, 山形 裕士

    日本植物生理学会2007年度年会, Mar. 2007, Japanese, 松山, Domestic conference

    Oral presentation

  • シロイヌナズナT7ファージ型RNAポリメラーゼRpoTp(RpoT;3)欠損株の解析

    稲垣 良作, 朴 杓允, 金地 通生, 塚谷 裕一, 柳田 一樹, 櫻井 望, 鈴木 秀幸, 柴田 大輔, 宇野 知秀, 山形 裕士, 金丸 研吾

    日本植物生理学会2007年度年会, Mar. 2007, Japanese, Domestic conference

    Oral presentation

  • シロイヌナズナgatABCホモローグ遺伝子の局在性と機能

    伊藤 滋一, 櫻井 望, 鈴木 秀幸, 柴田 大輔, 宇野 知秀, 山形 裕士, 金丸 研吾

    2007年度日本農芸化学会, Mar. 2007, Japanese, 東京, Domestic conference

    Oral presentation

  • ククミシン遺伝子の果実特異的発現機構の解析と果実の形質転換への応用

    奥山 慎也, 宇野 知秀, 金丸 研吾, 山形 裕士

    2007年度日本農芸化学会, Mar. 2007, Japanese, Domestic conference

    Oral presentation

  • ククミシンプロ配列の自己阻害活性に重要な領域

    中川 真隆, 宇野 知秀, 金丸 研吾, 山形 裕士

    2007年度日本農芸化学会, Mar. 2007, Japanese, 東京, Domestic conference

    Oral presentation

  • 葉緑体tRNAの動的な発現調節とその分子機構

    北村 元嗣, 柳田 一樹, 宇野 知秀, 山形 裕士, 金丸 研吾

    神大会館, Dec. 2006, Japanese, Domestic conference

    Poster presentation

  • 高等植物tRNAトランスアミデーション酵素の機能解析

    伊藤 滋一, 宇野 知秀, 山形 裕士, 金丸 研吾

    神戸大学研究基盤センター2006若手フロンティア研究会, Dec. 2006, Japanese, 神大会館, Domestic conference

    Poster presentation

  • ククミシンプロ配列の自己阻害に重要な領域

    中川 真隆, 上山 恵, 宇野 知秀, 金丸 研吾, 山形 裕士

    神戸大学研究基盤センター2006若手フロンティア研究会, Dec. 2006, Japanese, 神大会館, Domestic conference

    Poster presentation

  • Regulation of gene expression of soybean flavonoid-biosynthetic enzymes by cGMP

    吹田 憲治, 三井 麻衣子, 宇野 知秀, 金丸 研吾, 山形 裕士

    Kobe University Frontier Technology Forum, Nov. 2006, English, 神戸大学瀧川記念学術交流会館, Domestic conference

    Poster presentation

  • Molecular function of a T7 phage-type RNA polymerase in chloroplasts

    稲垣 良作, 伊藤 滋一, 一林 久雄, 柳田 一樹, 宇野 知秀, 山形 裕士, 金丸 研吾

    Kobe University Frontier Technology Forum, Nov. 2006, English, 神戸大学瀧川記念学術交流会館, Domestic conference

    Poster presentation

  • Basis and application of expression of two 5-ALA related enzymes in plants

    天尾 雅, 岩木 亜樹, 清田 真希, 高原 絵美, 宇野 知秀, 山形 裕士, 金丸 研吾

    Basis and application of expression of two 5’-ALA related enzymes in plants, Nov. 2006, English, 神戸大学瀧川記念学術交流会館, Domestic conference

    Poster presentation

  • Characterization of plastidic glutamyl-tRNA synthetase and ALA dehydratase in Arabidopsis.

    岩木 亜樹, 金地 通夫, 岡田 秀樹, 鈴木 秀幸, 柴田 大輔, 岩井 一弥, 鈴木 貴也, 宇野 知秀, 山形 裕士, 金丸 研吾

    11th IUPAC, Aug. 2006, English, 神戸, International conference

    Poster presentation

  • Pro-region of cucumisin precursor acts as a potent inhibitor of cucumisin

    中川 真隆, 上山 恵, 宇野 知秀, 金丸 研吾, 山形 裕士

    20th IUBMB International Congress of Biochemistry and Molecular Biology, Jun. 2006, English, IUBMB, 京都国際会館, International conference

    Poster presentation

  • Function of T7 Phage-Type RNA Polymerases in Arabidopsis (2) : RpoTmp (RpoT;2)

    Hisao Ichibayashi, 稲垣 良作, 朴 杓允, 金地 通生, 伊藤 滋一, 櫻井 望, 鈴木 秀幸, 柴田 大輔, 宇野 知秀, 山形 裕士, 金丸 研吾

    The 53rd NIBB Conference "Dynamic Organelles in Plant", Jun. 2006, English, Okazaki, International conference

    Poster presentation

  • Function of T7 Phage-Type RNA Polymerases in Arabidopsis (1) : RpoTp (RpoT;3)

    Inagaki Ryosaku, P. Pak, H. Tsukaya, Michio Kanechi, S. Itoh, Kazuki Ynagida, Nozomu Sakurai, Hideyuki Suzuki, Daisuke Shibata, 宇野 知秀, 山形 裕士, 金丸 研吾

    The 53rd NIBB Conference "Dynamic Organelles in Plant, Jun. 2006, English, Okazaki, International conference

    Poster presentation

  • Correlative and compensatory function of heterologous transcription systems in plastids

    金丸 研吾, 稲垣 良作, 一林 久雄, 宇野 知秀, 柴田 大輔, 山形 裕士

    20th IUBMB International Congress of Biochemistry and Molecular Biology, Jun. 2006, English, 京都, International conference

    Poster presentation

  • Cloning of cDNAs for Rab proteins from the brain of Bombyx mori and phosphorylation of their proteins expressed in Escherichia coli.

    中田 拓哉, 中尾 敦史, 山形 裕士, 金丸 研吾, 宇野 知秀

    20th IUBMB International Congress of Biochemistry and Molecular Biology, Jun. 2006, English, 京都, International conference

    Poster presentation

  • ダイズフラボノイド合成系酵素遺伝子の発現調節におけるcGMPとNOの機能相関

    Suita Kenji, Mitsui Maiko, Uno Tomohide, Kanamaru Kengo, Yamagata Hiroshi

    日本農芸化学会大会2006年度大会, Mar. 2006, Japanese, 京都, Domestic conference

    Poster presentation

  • シロイヌナズナT7ファージ型RNAポリメラーゼRpoT;2の機能解析

    Kengo Kanamaru, Ichibayashi Hisao, Inagaki Ryosaku, Ito Shigekazu, Kanachi Michio, Sakurai Nozomu, Suzuki Hideyuki, Shibata Daisuke, Uno Tomohide, Yamagata Hiroshi

    日本植物生理学会2006年度年会, Mar. 2006, Japanese, つくば, Domestic conference

    Oral presentation

  • ククミシンプロ配列の自己阻害活性

    Nakagawa Masataka, Ueyama Megumi, Uno Tomohide, Kanamaru Kengo, Yamagata Hiroshi

    日本農芸化学会大会2006年度大会, Mar. 2006, Japanese, 京都, Domestic conference

    Poster presentation

  • NO及びcGMPによって発現が調節されるシロイヌナズナ遺伝子の網羅的解析

    Ohara Tatsuya, Kiyota Maki, Ito Shigekazu, Inagaki Ryosaku, Suita Kenji, Sakurai Nozomu, Suzuki Hideyuki, Shibata Daisuke, Uno Tomohide, Kanamaru Kengo, Yamagata Hiroshi

    日本農芸化学会2006年度大会, Mar. 2006, Japanese, 京都, Domestic conference

    Poster presentation

  • シロイヌナズナT7ファージ型RNAポリメラーゼRpoT;2(RpoTmp)の機能解析

    一林 久雄, 稲垣 良作, 伊藤 滋一, 金地 通生, 櫻井 望, 鈴木 秀幸, 柴田 大輔, 宇野 知秀, 山形 裕士, 金丸 研吾

    第28回日本分子生物学会 講演要旨集p.449(f), Dec. 2005, Japanese, 日本分子生物学会, 福岡, Domestic conference

    Oral presentation

  • シロイヌナズナT87細胞の独立/混合条件における光応答の解析

    清田 真希, 関本 佳世子, 櫻井 望, 鈴木 秀幸, 柴田 大輔, 宇野 知秀, 山形 裕士, 金丸 研吾

    第78回日本生化学会大会 講演要旨集p.868(f), Oct. 2005, Japanese, 日本生化学会, 神戸, Domestic conference

    Oral presentation

  • ダイズELIP遺伝子の青色光/UV-A応答性発現を調節するシスエレメント

    野木 貴祐, 横山 輝之, 宇野 和秀, 金丸 研吾, 山形 裕士

    第12回日本光生物学協会年会 要旨集p.59(f), Aug. 2005, Japanese, 日本光生物学協会, 京都, Domestic conference

    Oral presentation

  • シロイヌナズナT87細胞の混合栄養培養と独立栄養培養における光応答の転写解析

    金丸 研吾, 清田 真希, 関本 佳世子, 櫻井 望, 鈴木 秀幸, 柴田 大輔, 宇野 知秀, 山形 裕士

    第23回日本植物分子細胞生物学会 p.193(f), Aug. 2005, Japanese, 日本植物分子細胞生物学会, 京都, Domestic conference

    Oral presentation

  • ダイズフラボノイド合成系遺伝子のcGMP およびNO による発現調節

    吹田 憲治, 三井 麻衣子, 宇野 知秀, 金丸 研吾, 山形 裕士

    第52回日本生化学会近畿支部例会 要旨集p.38(f), May 2005, Japanese, 日本生化学会近畿支部, 神戸大学, Domestic conference

    Oral presentation

  • カイコのメチル化DNA 結合タンパク質の分子特性

    野村 由佳, 岩永 陽介, 中尾 淳史, 中村 正彦, 田嶋 正二, 金丸 研吾, 山形 裕士, 宇野 知秀

    第52回日本生化学会近畿支部例会 要旨集p.37(f), May 2005, Japanese, 日本生化学会近畿支部, 神戸大学, Domestic conference

    Oral presentation

  • 昆虫の脳に存在する蛋白質のPKCによるリン酸化およびその機能解析

    中田 拓哉, 中尾 淳史, 金丸 研吾, 山形 裕士, 宇野 知秀

    第75回蚕糸昆虫機能学術講演会, Apr. 2005, Japanese, 日本蚕糸学会, 東京, Domestic conference

    Oral presentation

  • 1原子酸素添加酵素チトクロームP450 を用いたバイオコンビケムへの応用

    増田 智子, 中尾 淳史, 宇野 知秀, 谷口 雄規, 山形 裕士, 金丸 研吾, 大野 清治, 今石 浩正

    第20回Combinatorial Chemistry研究会, Apr. 2005, Japanese, Combinatorial Chemistry研究会, 大阪, Domestic conference

    Oral presentation

  • 果実特異的遺伝子発現を調節するエンハンサー因子のcDNA クローニングと特性解析

    東 礼奈, 柴本 憲幸, 宇野 知秀, 金丸 研吾, 山形 裕士

    日本農芸化学会大会2005年度大会 要旨集p.150, Mar. 2005, Japanese, 日本農芸化学会, 札幌, Domestic conference

    Oral presentation

  • ダイズフラボノイド合成系酵素遺伝子のcGMPおよび光による発現調節

    吹田 憲治, 三井 麻衣子, 宇野 知秀, 金丸 研吾, 山形 裕士

    日本農芸化学会大会2005年度大会 要旨集p.149(f), Mar. 2005, Japanese, 日本農芸化学会, 札幌, Domestic conference

    Oral presentation

  • シロイヌナズナ葉緑体tRNA の発現ダイナミクス

    柳田 一樹, 宇野 知秀, 金丸 研吾, 山形 裕士

    日本農芸化学会大会2005年度大会 要旨集p.150(f), Mar. 2005, Japanese, 日本農芸化学会, 札幌, Domestic conference

    Oral presentation

  • イネ種子二機能性酵素インヒビター(RASI)の大腸菌における発現およびα� アミラーゼ阻害特性

    出口 正揮, 宇野 知秀, 金丸 研吾, 山形 裕士

    日本農芸化学会大会2005年度大会 要旨集p.311(f), Mar. 2005, Japanese, 日本農芸化学会, 札幌, Domestic conference

    Oral presentation

  • 1原子酸素添加酵素チトクロームP450 を用いたバイオコンビケムへの応用

    増田 智子, 中尾 敦史, 宇野 知秀, 谷口 雄規, 山形 裕士, 金丸 研吾, 大野 清春, 今石 浩正

    第21回Combinatorial Chemistry 研究会, 2005, Japanese, Combinatorial Chemistry 研究会, 東京, Domestic conference

    Oral presentation

  • 高等植物葉緑体におけるtRNA発現の重要性とそのダイナミクス

    柳田 一樹, 宇野 知秀, 山形 裕士, 金丸 研吾

    日本農芸化学会関西支部第437回講演会), 2004, Japanese, 日本農芸化学会関西支部, 未記入, Domestic conference

    Oral presentation

  • ダイズELIP遺伝子プロモーターの青色光/紫外光に対する光応答能

    横山 輝之, 宇野 知秀, 金丸 研吾, 山形 裕士

    日本農芸化学会2004年度大会, 2004, Japanese, 日本農芸化学会, 未記入, Domestic conference

    Oral presentation

  • イネ種子二機能性酵素インヒビター(RASI)の大腸菌における発現およびα-アミラーゼ阻害特性

    出口 正輝, 宇野 知秀, 金丸 研吾, 山形 裕士

    第25回種子生理生化学研究会, 2004, Japanese, 未記入, 未記入, Domestic conference

    Oral presentation

  • ダイズフラボノイド合成系酵素遺伝子のcGMP による発現調節

    吹田 憲治, 三井 麻衣子, 宇野 知秀, 金丸 研吾, 山形 裕士

    日本農芸化学会関西支部第432回講演会, Dec. 2003, Japanese, 日本農芸化学会関西支部, 神戸大学, Domestic conference

    Oral presentation

  • カイコ脳低分子量G 蛋白質(Rab)のcDNA クローニングとその発現

    宇野 知秀, 平垣 進

    日本蚕糸学会第73回大会講演要旨集, Mar. 2003, Japanese, 日本蚕糸学会, 東京, Domestic conference

    Oral presentation

  • Small GTP binding proteins from the brain of Bombyx mori.

    NAKAO A, KATSURAUMA C, UNO Tomohide, KANAMARU Kengo, YAMAGATA Hiroshi

    Japan-Korea Joint Seminar on Sericultural Sciences and Insect Industry, 2003, English, 未記入, 未記入, International conference

    Oral presentation

  • Phosphatase I(低温酵素)の分子特性

    Hiroki Tsuruta, Tomohide Uno, Keiichi Watanabe, Yasuo Aizono

    日本農芸化学会西日本・関西合同大会, 1997, Japanese, 日本農芸化学会西日本・関西支部, 佐賀, Domestic conference

    Oral presentation

Association Memberships

  • 日本蚕糸学会

Research Projects

  • 宇野 知秀

    学術研究助成基金助成金/基盤研究(C), Apr. 2014 - Mar. 2017, Principal investigator

    Competitive research funding

  • 身近にある食品や生物の蛋白質や遺伝子を調べよう。

    宇野 知秀

    独立行政法人日本学術振興会, 研究成果の社会還元・普及事業 ひらめき☆ときめきサイエンス, 2017, Principal investigator

    Competitive research funding

  • 生物の蛋白質と遺伝子を取り出し、色々と調べよう

    宇野 知秀

    ひらめき☆ときめきサイエンス, 2014, Principal investigator

    Competitive research funding

  • 身近なものの蛋白質と遺伝子をみてみよう

    宇野 知秀

    ひらめき☆ときめきサイエンス, 2013, Principal investigator

    Competitive research funding

  • 色々な生物の蛋白質と遺伝子を調べてみよう

    宇野知秀

    ひらめき☆ときめきサイエンス, 2012, Principal investigator

    Competitive research funding

  • 昆虫の蛋白質を見てみよう-昆虫を用いた生化学実験-

    宇野 知秀

    ひらめき☆ときめきサイエンス, 2011, Principal investigator

    Competitive research funding

  • 宇野 知秀

    科学研究費補助金/基盤研究(C), 2008, Principal investigator

    Competitive research funding

  • 宇野 知秀

    科学研究費補助金/基盤研究(C), 2006, Principal investigator

    Competitive research funding

  • 昆虫の脳特異的低分子量GTP結合蛋白質の機能特性

    宇野 知秀

    日本学術振興会, 科学研究費助成事業, 若手研究(B), 神戸大学, 2002 - 2003

    低分子量GTP結合蛋白質(s-GBP)は、多数のスーパーファミリーを形成するが、その内Rab蛋白質は、神経ペプチドや、蛋白質の輸送に関与する。一方、我々は阻害剤を用いた研究により、昆虫の神経ペプチドであるPTTH(Prothoracicotropic hormone)の分泌にs-GBPのリン酸化が関与することを明らかにした。 本研究では、昆虫の神経ペプチド分泌に関与するs-GBPが果たす役割を分子レベルで明らかにするために、s-GBPをリン酸化するprotein kinase及びリン酸化されたs-GBPの分子機能を調べる。 そこで、まずカイコの脳より3種類の全長cDNAを含むRab蛋白質(Rab1,Rab8,Rab14)を発現した。この内Rab14については、ratの脳のprotein kinaseによりリン酸化されることを明らかにした(Arch Insect Biochem Physiol,2003)。一方カイコ脳より各種カラムクロマトグラフィーを用いて、各種Protein Kinaseを分離した[Protein kinase C (PKC),Ca/CaM dependent protein kinase (CaM kinase),Nucleoside diphosphate kinase(NDP kinase)]。その内NDP kinaseについては、完全に精製することができた(Arch Insect Biochem Physiol,2002)。これらのkinaseを用いてRab蛋白質をリン酸化したところ、Rab8とRab14は、CaM kinaseによりわずかに、そして、PKCにより、強くリン酸化されることがわかった(Arch Insect Biochem, Physiol,投稿中)。現在、このリン酸化されたRab蛋白質の分子特性を調べている。また、Rabをリン酸化するPKCについて、その分子種を明らかにするために、PKCの2種類のcDNAを動物細胞で発現させた。これらの発現蛋白質はProtein Kinase活性を有していた(投稿準備中)。現在この発現蛋白質がRab蛋白質をリン酸化するか調べている。

  • 低温酵素(Psychrophilic enzyme)の分子機能に関する研究

    相薗 泰生, 宇野 知秀

    日本学術振興会, 科学研究費助成事業, 萌芽的研究, 神戸大学, 1997 - 1998

    低温域(0〜5°C)で作用する酵素の触媒機能を高次構造との関連で解明する研究の一環として、好冷菌(Shewanella sp.)が産生するアルカリフォスファターゼに着目して、その分離精製を行い、精製酵素について酵素化学的及び理化学的性質を明らかにした。 ●アルカリフォスファターゼ(AP)の分離精製:菌体(2Kg)を磨砕して得られた酵素抽出液を、硫酸アンモニウム40-80%飽和で粗分画した後、順次、DEAE-ce11ulofine、Butyl-cellulofine、Hydroxylapatite、Sephacryl S-100、Mono-Qを用いたカラムクロマトグラフィーに供し、Native-PAGE及びSDS-PAGEで均一な酵素標品1.04mg(比活性:1394)を活性収率13.1%で分離した。●APの酵素化学的特性:精製酵素は、至適pH9.8、至適温度40℃を示し、pH9.8、温度0〜10℃の範囲で安定であったが、20℃以上で失活する特性を示した。本酵素は、金属イオンを要求し、Mg^<2+>、Mn^<2+>及びCo^<2+>により活性が高められた。0℃における活性は、40℃(至適温度)の39%を示した。また、幅広い基質特異性を示し、ATP、GTP、CTP、β-Glycerophosphateなどを脱リン酸化した。●APの理化学的特性:SDS-PAGE及びアミノ酸組成分析で測定された分子量は、41,800であった。また、精密アミノ酸組成分析により、この酵素は、高次構造の安定化因子であるプロリン残基の含量が低く、また、分子内に1つのジスルフィド結合を有していることが明らかになった。これらの特性が、この酵素の中温域における不安定性と低温域における高い触媒活性に寄与しているものと推測された。 ●Phosphatase Iの酵素化学的特性:昨年、同じ菌株から単離したPhosphatase Iについて、基質特異性と、また化学修飾により触媒残基を詳細に検討し、本酵素は、Hisを触媒残基とするProtein-tyrosine-phosphataseであることを明らかにした。

Industrial Property Rights

  • 膜結合型P450発現用カセットプラスミド

    IMAISHI HIROMASA, UNO TOMOHIDE

    特願2006-132484, 11 May 2006, 大学長, 特許4899050, 13 Jan. 2012

    Patent right