Research activity information

• Apr. 2023 長瀬研究振興賞


• 2020 日本糖質学会, 日本糖質学会奨励賞, 高等動物におけるN 型糖鎖のマンノース切除を基軸とした小胞体関連分解機構の解析


• 2020 日本生化学会, 日本生化学会奨励賞, 遺伝子破壊法を用いた小胞体関連分解機構の解明


• 2017 日本細胞生物学会, CSF award, SEL1L-dependent Substrates Require Derlin2/3 and Herp1/2 for Endoplasmic Reticulum-associated Degradation

  杉本 岳大、蜷川 暁


• Jun. 2016 日本細胞生物学会, 若手優秀発表賞, 分解執行局域における新展開


• 2008 Kyoto University Evolution and Biodiversity from Genome to Ecosystem, gCOE poster Award, Identification and analysis of Derlins Interacting Protein

• Systematic and comprehensive analysis of major localizations of alpha-dystroglycan-specific modifying enzymes

  Shinya Aso, Martin Lowe, Kazutoshi Mori, Satoshi Ninagawa

  Jun. 2025, Glycobiology

  Scientific journal


• UGGT1-mediated reglucosylation of N-glycan competes with ER-associated degradation of unstable and misfolded glycoproteins

  Satoshi Ninagawa, Masaki Matsuo, Deng Ying, Shuichiro Oshita, Shinya Aso, Kazutoshi Matsushita, Mai Taniguchi, Akane Fueki, Moe Yamashiro, Kaoru Sugasawa, Shunsuke Saito, Koshi Imami, Yasuhiko Kizuka, Tetsushi Sakuma, Takashi Yamamoto, Hirokazu Yagi, Koichi Kato, Kazutoshi Mori

  Dec. 2024, eLife

  Scientific journal

  Link to Kobe Univ. Repository (Kernel)


• Ca²⁺-driven PDIA6 phase separation to ensure proinsulin quality control

  Young-Ho Lee, Tomohide Saio, Mai Watabe, Motonori Matsusaki, Shingo Kanemura, Yuxi Lin, Taro Mannen, Tsubura Kuramochi, Katsuya Iuchi, Michiko Tajiri, Kotono Suzuki, Yan Li, Yunseok Heo, Yuka Kamada, Kenta Arai, Mayuko Hashimoto, Satoshi Ninagawa, Yoshikazu Hattori, Hiroyuki Kumeta, Airu Takeuchi, Hiroya Abe, Eiichiro Mori, Takahiro Muraoka, Tsukasa Okiyoneda, Satoko Akashi, Michele Vendruscolo, Kenji Inaba, Masaki Okumura

  Abstract The endoplasmic reticulum (ER) plays key roles in protein quality control^1,2and dynamic Ca²⁺storage^3,4in eukaryotic cells. However, the protein homeostasis (proteostasis) system that regulates these ER functions is still incompletely characterised. Previous study revealed the importance of oligomerization in the function PDIA1, an ER-resident disulfide isomerase and molecular chaperone, regulates oligomeric states in accordance with client folding⁵. This result suggests that at least some of the 20 members of other PDI family may undergo regulated self-assembly in order to optimally function. Here, we show that Ca²⁺triggers the phase separation of PDIA6 into liquid-like condensates. In contrast to the condensation mechanism observed for proteins containing low-complexity domains, our results indicate that transient but specific electrostatic interactions occur between the first and the third folded thioredoxin-like domains of PDIA6. We further show that the Ca²⁺-driven condensation of PDIA6 recruits PDIA3 and proinsulin, thus increasing their local concentrations. This process results in the 30-fold enhancement of proinsulin folding and in the inhibition of proinsulin aggregation. Our findings shed light on a condensation-driven Ca²⁺-mediated proteostasis cascade in the ER by revealing how the efficiency of the protein folding process can be enhanced within quality control granules.

  Cold Spring Harbor Laboratory, Jul. 2024


• Proteomic analysis of fatty liver induced by starvation of medaka fish larvae.

  Tomoyo Ikeda, Tokiro Ishikawa, Satoshi Ninagawa, Tetsuya Okada, Masaya Ono, Kazutoshi Mori

  When medaka fish (Oryzias latipes) larvae are grown in the absence of exogenous nutrition, the liver becomes dark and positive to Oil Red O staining from 7 days post-hatch (dph). We determined the mechanism of this starvation-induced development of fatty liver by proteomic analysis using livers obtained from larvae grown in the presence or absence of 2% glucose at 5 dph. Results showed that changes in the expression levels of enzymes involved in glycolysis or the tricarboxylic acid cycle were modest, whereas the expression levels of enzymes involved in amino acid catabolism or β-oxidation of fatty acids were significantly elevated, suggesting that they become major energy sources under starvation conditions. Expression levels of enzymes for the uptake and β-oxidation of fatty acids as well as synthesis of triacylglycerol were elevated, whereas those for the synthesis of cholesterol as well as export of cholesterol and triacylglycerol were decreased under starvation conditions, which explains the accumulation of triacylglycerol in the liver. Our results provide the basis for future research to understand how gene malfunction(s) affects the development of fatty liver, which can lead to nonalcoholic steatohepatitis and then to liver cirrhosis.Key words: amino acid catabolism, β-oxidation, triacylglycerol, cholesterol, export.

  Jul. 2023, Cell structure and function, 48(2) (2), 123 - 133, English, Domestic magazine

  Scientific journal


• Loss of ATF6α in a Human Carcinoma Cell Line Is Compensated not by Its Paralogue ATF6β but by Sustained Activation of the IRE1 and PERK Arms for Tumor Growth in Nude Mice

  Shengyu Jin, Byungseok Jin, Tokiro Ishikawa, Satoshi Ninagawa, Tetsuya Okada, Sho Koyasu, Hiroshi Harada, Kazutoshi Mori

  To survive poor nutritional conditions, tumor cells activate the unfolded protein response, which is composed of the IRE1, PERK and ATF6 arms, to maintain the homeostasis of the endoplasmic reticulum, where secretory and transmembrane proteins destined for the secretory pathway gain their correct three dimensional structure. The requirement of the IRE1 and PERK arms for tumor growth in nude mice is established. Here, we investigated the requirement for the ATF6 arm, which consists of ubiquitously expressed ATF6α and ATF6β, by constructing ATF6α-knockout, ATF6β-knockout and ATF6α/β-double knockout in HCT116 cells derived from human colorectal carcinoma. Results showed that these knockout cells grew similarly to wild-type cells in nude mice, contrary to expectations from our analysis of ATF6α-knockout, ATF6β-knockout and ATF6α/β-double knockout mice. We then found that the loss of ATF6α in HCT116 cells resulted in sustained activation of the IRE1 and PERK arms, in marked contrast to mouse embryonic fibroblasts, in which the loss of ATF6α is compensated for by ATF6β. Although IRE1-knockout in HCT116 cells unexpectedly did not affect tumor growth in nude mice, IRE1-knockout HCT116 cells with ATF6α knockdown grew significantly more slowly than wild-type or IRE1-knockout HCT116 cells. These results have unraveled the situation-dependent differential compensation strategies of ATF6α.

  American Society for Cell Biology (ASCB), Jan. 2023, Molecular Biology of the Cell

  Scientific journal


• A motor neuron disease-associated mutation produces non-glycosylated Seipin that induces ER stress and apoptosis by inactivating SERCA2b

  Shunsuke Saito, Tokiro Ishikawa, Satoshi Ninagawa, Tetsuya Okada, Kazutoshi Mori

  A causal relationship between endoplasmic reticulum (ER) stress and the development of neurodegenerative diseases remains controversial. Here, we focused on Seipinopathy, a dominant motor neuron disease, based on the finding that its causal gene product, Seipin, is a protein that spans the ER membrane twice. Gain-of-function mutations of Seipin produce non-glycosylated Seipin (ngSeipin), which was previously shown to induce ER stress and apoptosis at both cell and mouse levels albeit with no clarified mechanism. We found that aggregation-prone ngSeipin dominantly inactivated SERCA2b, the major calcium pump in the ER, and decreased the calcium concentration in the ER, leading to ER stress and apoptosis in human colorectal carcinoma-derived cells (HCT116). This inactivation required oligomerization of ngSeipin and direct interaction of the C-terminus of ngSeipin with SERCA2b, and was observed in Seipin-deficient neuroblastoma (SH-SY5Y) cells expressing ngSeipin at an endogenous protein level. Our results thus provide a new direction to the controversy noted above.

  Nov. 2022, eLife, 11, English, International magazine

  Scientific journal


• Purified EDEM3 or EDEM1 alone produces determinant oligosaccharide structures from M8B in mammalian glycoprotein ERAD.

  Ginto George, Satoshi Ninagawa, Hirokazu Yagi, Jun-Ichi Furukawa, Noritaka Hashii, Akiko Ishii-Watabe, Ying Deng, Kazutoshi Matsushita, Tokiro Ishikawa, Yugoviandi P Mamahit, Yuta Maki, Yasuhiro Kajihara, Koichi Kato, Tetsuya Okada, Kazutoshi Mori

  Sequential mannose trimming of N-glycan, from M9 to M8B and then to oligosaccharides exposing the α1,6-linked mannosyl residue (M7A, M6, and M5), facilitates endoplasmic reticulum-associated degradation of misfolded glycoproteins (gpERAD). We previously showed that EDEM2 stably disulfide-bonded to the thioredoxin domain-containing protein TXNDC11 is responsible for the first step (George et al., 2020). Here, we show that EDEM3 and EDEM1 are responsible for the second step. Incubation of pyridylamine-labeled M8B with purified EDEM3 alone produced M7 (M7A and M7C), M6, and M5. EDEM1 showed a similar tendency, although much lower amounts of M6 and M5 were produced. Thus, EDEM3 is a major α1,2-mannosidase for the second step from M8B. Both EDEM3 and EDEM1 trimmed M8B from a glycoprotein efficiently. Our confirmation of the Golgi localization of MAN1B indicates that no other α1,2-mannosidase is required for gpERAD. Accordingly, we have established the entire route of oligosaccharide processing and the enzymes responsible.

  Lead, Oct. 2021, eLife, 10, English, International magazine

  [Refereed]

  Scientific journal

  Link to Kobe Univ. Repository (Kernel)


• Mechanisms of productive folding and endoplasmic reticulum-associated degradation of glycoproteins and non-glycoproteins.

  Satoshi Ninagawa, Ginto George, Kazutoshi Mori

  BACKGROUND: The quality of proteins destined for the secretory pathway is ensured by two distinct mechanisms in the endoplasmic reticulum (ER): productive folding of newly synthesized proteins, which is assisted by ER-localized molecular chaperones and in most cases also by disulfide bond formation and transfer of an oligosaccharide unit; and ER-associated degradation (ERAD), in which proteins unfolded or misfolded in the ER are recognized and processed for delivery to the ER membrane complex, retrotranslocated through the complex with simultaneous ubiquitination, extracted by AAA-ATPase to the cytosol, and finally degraded by the proteasome. SCOPE OF REVIEW: We describe the mechanisms of productive folding and ERAD, with particular attention to glycoproteins versus non-glycoproteins, and to yeast versus mammalian systems. MAJOR CONCLUSION: Molecular mechanisms of the productive folding of glycoproteins and non-glycoproteins mediated by molecular chaperones and protein disulfide isomerases are well conserved from yeast to mammals. Additionally, mammals have gained an oligosaccharide structure-dependent folding cycle for glycoproteins. The molecular mechanisms of ERAD are also well conserved from yeast to mammals, but redundant expression of yeast orthologues in mammals has been encountered, particularly for components involved in recognition and processing of glycoproteins and components of the ER membrane complex involved in retrotranslocation and simultaneous ubiquitination of glycoproteins and non-glycoproteins. This may reflect an evolutionary consequence of increasing quantity or quality needs toward mammals. GENERAL SIGNIFICANCE: The introduction of innovative genome editing technology into analysis of the mechanisms of mammalian ERAD, as exemplified here, will provide new insights into the pathogenesis of various diseases.

  Dec. 2020, Biochimica et biophysica acta. General subjects, 129812 - 129812, English, International magazine

  Scientific journal


• Antipsychotic olanzapine-induced misfolding of proinsulin in the endoplasmic reticulum accounts for atypical development of diabetes.

  Satoshi Ninagawa, Seiichiro Tada, Masaki Okumura, Kenta Inoguchi, Misaki Kinoshita, Shingo Kanemura, Koshi Imami, Hajime Umezawa, Tokiro Ishikawa, Robert B Mackin, Seiji Torii, Yasushi Ishihama, Kenji Inaba, Takayuki Anazawa, Takahiko Nagamine, Kazutoshi Mori

  Second-generation antipsychotics are widely used to medicate patients with schizophrenia, but may cause metabolic side effects such as diabetes, which has been considered to result from obesity-associated insulin resistance. Olanzapine is particularly well known for this effect. However, clinical studies have suggested that olanzapine-induced hyperglycemia in certain patients cannot be explained by such a generalized mechanism. Here, we focused on the effects of olanzapine on insulin biosynthesis and secretion by mouse insulinoma MIN6 cells. Olanzapine reduced maturation of proinsulin, and thereby inhibited secretion of insulin; and specifically shifted the primary localization of proinsulin from insulin granules to the endoplasmic reticulum. This was due to olanzapine's impairment of proper disulfide bond formation in proinsulin, although direct targets of olanzapine remain undetermined. Olanzapine-induced proinsulin misfolding and subsequent decrease also occurred at the mouse level. This mechanism of olanzapine-induced β-cell dysfunction should be considered, together with weight gain, when patients are administered olanzapine.

  Nov. 2020, eLife, 9, e60970, English, International magazine

  [Refereed]

  Scientific journal


• Improved secretion of glycoproteins using an N-glycan-restricted passport sequence tag recognized by cargo receptor.

  Hirokazu Yagi, Maho Yagi-Utsumi, Rena Honda, Yusaku Ohta, Taiki Saito, Miho Nishio, Satoshi Ninagawa, Kousuke Suzuki, Takahiro Anzai, Yukiko Kamiya, Kazuhiro Aoki, Mahito Nakanishi, Tadashi Satoh, Koichi Kato

  MCFD2 and ERGIC-53, which are the products of causative genes of combined factor V and factor VIII deficiency, form a cargo receptor complex responsible for intracellular transport of these coagulation factors in the early secretory pathway. In this study, using an NMR technique, we successfully identified an MCFD2-binding segment from factor VIII composed of a 10 amino acid sequence that enhances its secretion. This prompted us to examine possible effects of attaching this sequence to recombinant glycoproteins on their secretion. We found that the secretion level of recombinant erythropoietin was significantly increased simply by tagging it with the passport sequence. Our findings not only provide molecular basis for the intracellular trafficking of coagulation factors and their genetic deficiency but also offer a potentially useful tool for increasing the production yields of recombinant glycoproteins of biopharmaceutical interest.

  Mar. 2020, Nature communications, 11(1) (1), 1368 - 1368, English, International magazine

  [Refereed]

  Scientific journal


• EDEM2 stably disulfide-bonded to TXNDC11 catalyzes the first mannose trimming step in mammalian glycoprotein ERAD.

  Ginto George, Satoshi Ninagawa, Hirokazu Yagi, Taiki Saito, Tokiro Ishikawa, Tetsushi Sakuma, Takashi Yamamoto, Koshi Imami, Yasushi Ishihama, Koichi Kato, Tetsuya Okada, Kazutoshi Mori

  Sequential mannose trimming of N-glycan (Man9GlcNAc2 -> Man8GlcNAc2 -> Man7GlcNAc2) facilitates endoplasmic reticulum-associated degradation of misfolded glycoproteins (gpERAD). Our gene knockout experiments in human HCT116 cells have revealed that EDEM2 is required for the first step. However, it was previously shown that purified EDEM2 exhibited no α1,2-mannosidase activity toward Man9GlcNAc2 in vitro. Here, we found that EDEM2 was stably disulfide-bonded to TXNDC11, an endoplasmic reticulum protein containing five thioredoxin (Trx)-like domains. C558 present outside of the mannosidase homology domain of EDEM2 was linked to C692 in Trx5, which solely contains the CXXC motif in TXNDC11. This covalent bonding was essential for mannose trimming and subsequent gpERAD in HCT116 cells. Furthermore, EDEM2-TXNDC11 complex purified from transfected HCT116 cells converted Man9GlcNAc2 to Man8GlcNAc2(isomerB) in vitro. Our results establish the role of EDEM2 as an initiator of gpERAD, and represent the first clear demonstration of in vitro mannosidase activity of EDEM family proteins.

  Lead, Feb. 2020, eLife, 9, e53455, English, International magazine

  [Refereed]

  Scientific journal


• Development of a Rapid in vivo Assay to Evaluate the Efficacy of IRE1-specific Inhibitors of the Unfolded Protein Response Using Medaka Fish.

  Byungseok Jin, Tokiro Ishikawa, Mai Taniguchi, Satoshi Ninagawa, Tetsuya Okada, Shigehide Kagaya, Kazutoshi Mori

  Three types of transmembrane protein, IRE1α/IRE1β, PERK, and ATF6α/ATF6β, are expressed ubiquitously in vertebrates as transducers of the unfolded protein response (UPR), which maintains the homeostasis of the endoplasmic reticulum. IRE1 is highly conserved from yeast to mammals, and transmits a signal by a unique mechanism, namely splicing of mRNA encoding XBP1, the transcription factor downstream of IRE1 in metazoans. IRE1 contains a ribonuclease domain in its cytoplasmic region which initiates splicing reaction by direct cleavage of XBP1 mRNA at the two stem loop structures. As the UPR is considered to be involved in the development and progression of various diseases, as well as in the survival and growth of tumor cells, UPR inhibitors have been sought. To date, IRE1 inhibitors have been screened using cell-based reporter assays and fluorescent-based in vitro cleavage assays. Here, we used medaka fish to develop an in vivo assay for IRE1α inhibitors. IRE1α, IRE1β, ATF6α and ATF6β are ubiquitously expressed in medaka. We found that IRE1α/ATF6α-double knockout is lethal, similarly to IRE1α/IRE1β- and ATF6α/ATF6β-double knockout. Therefore, IRE1 inhibitors are expected to confer lethality to ATF6α-knockout medaka but not to wild-type medaka. One compound named K114 was obtained from 1,280 compounds using this phenotypic screening. K114 inhibited ER stress-induced splicing of XBP1 mRNA as well as reporter luciferase expression in HCT116 cells derived from human colorectal carcinoma, and inhibited ribonuclease activity of human IRE1α in vitro. Thus, this phenotypic assay can be used as a quick test for the efficacy of IRE1α inhibitors in vivo.Key words: endoplasmic reticulum, inhibitor screening, mRNA splicing, phenotypic assay, unfolded protein response.

  Feb. 2020, Cell structure and function, 45(1) (1), 23 - 31, English, Domestic magazine

  [Refereed]

  Scientific journal


• Reinvestigation of Disulfide-bonded Oligomeric Forms of the Unfolded Protein Response Transducer ATF6.

  Hibiki Koba, Shengyu Jin, Nanami Imada, Tokiro Ishikawa, Satoshi Ninagawa, Tetsuya Okada, Tetsushi Sakuma, Takashi Yamamoto, Kazutoshi Mori

  ATF6α is an endoplasmic reticulum (ER)-embedded transcription factor which is rapidly activated by ER stress, and a major regulator of ER chaperone levels in vertebrates. We previously suggested that ATF6α occurs as a monomer, dimer and oligomer in the unstressed ER of Chinese hamster ovary cells due to the presence of two evolutionarily conserved cysteine residues in its luminal region (C467 and C618), and showed that ATF6α is reduced upon ER stress, such that only reduced monomer ATF6α is translocated to the Golgi apparatus for activation by proteolysis. However, mutagenesis analysis (C467A and C618A) revealed that the C618A mutant behaves in an unexpected manner (monomer and oligomer) during non-reducing SDS-PAGE, for reasons which remained unclear. Here, we used human colorectal carcinoma-derived HCT116 cells deficient in ATF6α and its relevant ATF6β, and found that ATF6α dimer and oligomer are both dimers, which we designated C618-dimer and C467-dimer, respectively. We demonstrated that C467-dimer (previously considered an oligomer) behaved bigger than C618-dimer (previously considered a dimer) during non-reducing SDS-PAGE, based on their disulfide-bonded structures. Furthermore, ATF6α monomer physically associates with another ATF6α monomer in the absence of disulfide bonding, which renders two C467 residues in close proximity so that formation of C467-dimer is much easier than that of C618-dimer. In contrast, C618-dimer is more easily reduced upon ER stress. Thus, our analysis revealed that all forms of ATF6α, namely monomer, C618-dimer and C467-dimer, are activated by single reduction of a disulfide bond in response to ER stress, ensuring the rapidity of ATF6α activation.Key words: disulfide-bonded structure, endoplasmic reticulum, membrane-bound transcription factor, non-reducing SDS-PAGE, unfolded protein response.

  Jan. 2020, Cell structure and function, 45(1) (1), 9 - 21, English, Domestic magazine

  [Refereed]

  Scientific journal


• メダカを用いた小胞体ストレスに起因するアポトーシス分子機構の解析

  陳 炳碩, 石川 時郎, 蜷川 暁, 岡田 徹也, 森 和俊

  (公社)日本生化学会, Sep. 2019, 日本生化学会大会プログラム・講演要旨集, 92回, [1T05m - 01], Japanese


• 小胞体ストレスセンサーATF6αはジスルフィド結合を介して2種類の二量体を形成する(Endoplasmic reticulum stress sensor ATF6α forms two types of dimers via disulfide-bonding)

  古場 玲, 金 聖宇, 岡田 徹也, 石川 時郎, 蜷川 暁, 森 和俊

  (公社)日本生化学会, Sep. 2019, 日本生化学会大会プログラム・講演要旨集, 92回, [3T03m - 01], Japanese


• 小胞体膜タンパク質Seipinの神経変性誘導変異体に起因する小胞体ストレス発生機構の解析

  齊藤 峻介, 石川 時郎, 蜷川 暁, 岡田 徹也, 森 和俊

  (公社)日本生化学会, Sep. 2019, 日本生化学会大会プログラム・講演要旨集, 92回, [3T17a - 03], Japanese


• 第二世代抗精神病薬オランザピンが、副作用として糖尿病を引き起こす分子メカニズムの解析 インスリンは、適切なジスルフィド結合形成を阻害され、小胞体関連分解により処理される

  蜷川 暁, 岡田 徹也, 梅澤 元, 石川 時郎, 鳥居 征司, Mackin Robert, 長嶺 敬彦, 森 和俊

  生命科学系学会合同年次大会運営事務局, Dec. 2017, 生命科学系学会合同年次大会, 2017年度, [1P - 1141], Japanese


• SEL1L-dependent Substrates Require Derlin2/3 and Herp1/2 for Endoplasmic Reticulum-associated Degradation.

  Takehiro Sugimoto, Satoshi Ninagawa, Shimpei Yamano, Tokiro Ishikawa, Tetsuya Okada, Shunichi Takeda, Kazutoshi Mori

  Lead, Jul. 2017, Cell structure and function, 42(2) (2), 81 - 94, English, Domestic magazine

  [Refereed]

  Scientific journal


• UPR transducer BBF2H7 allows export of type II collagen in a cargo- and developmental stage-specific manner.

  Tokiro Ishikawa, Takuya Toyama, Yuki Nakamura, Kentaro Tamada, Hitomi Shimizu, Satoshi Ninagawa, Tetsuya Okada, Yasuhiro Kamei, Tomoko Ishikawa-Fujiwara, Takeshi Todo, Eriko Aoyama, Masaharu Takigawa, Akihiro Harada, Kazutoshi Mori

  Jun. 2017, The Journal of cell biology, 216(6) (6), 1761 - 1774, English, International magazine

  [Refereed]

  Scientific journal


• Direct Mapping of Additional Modifications on Phosphorylated O-glycans of α-Dystroglycan by Mass Spectrometry Analysis in Conjunction with Knocking Out of Causative Genes for Dystroglycanopathy.

  Hirokazu Yagi, Chu-Wei Kuo, Takayuki Obayashi, Satoshi Ninagawa, Kay-Hooi Khoo, Koichi Kato

  Dystroglycanopathy is a major class of congenital muscular dystrophy caused by a deficiency of functional glycans on α-dystroglycan (αDG) with laminin-binding activity. Recent advances have led to identification of several causative gene products of dystroglycanopathy and characterization of their in vitro enzymatic activities. However, the in vivo functional roles remain equivocal for enzymes such as ISPD, FKTN, FKRP, and TMEM5 that are supposed to be involved in post-phosphoryl modifications linking the GalNAc-β3-GlcNAc-β4-Man-6-phosphate core and the outer laminin-binding glycans. Herein, by direct nano-LC-MS2/MS3 analysis of tryptic glycopeptides derived from a truncated recombinant αDG expressed in the wild-type and a panel of mutated cells deficient in one of these enzymes, we sought to define the full extent of variable modifications on this phosphorylated core O-glycan at the functional Thr317/Thr319 sites. We showed that the most abundant glycoforms carried a phosphorylated core at each of the two sites, with and without a single ribitol phosphate (RboP) extending from terminal HexNAc. At much lower signal intensity, a novel substituent tentatively assigned as glycerol phosphate (GroP) was additionally detected. As expected, tandem RboP extended with a GlcA-Xyl unit was only identified in wild type, whereas knocking out of either ISPD or FKTN prevented formation of RboP. In the absence of FKRP, glycoforms with single but not tandem RboP accumulated, consistent with the suggested role of this enzyme in transferring the second RboP. Intriguingly, the single GroP modification also required functional FKTN whereas absence of TMEM5 significantly hindered only the addition of RboP. Our findings thus revealed additional levels of complexity associated with the core structures, suggesting functional interplay among these enzymes through their interactions. The simplified analytical workflow developed here should facilitate rapid mapping across a wider range of cell types to gain better insights into its physiological relevance.

  Nov. 2016, Molecular & cellular proteomics : MCP, 15(11) (11), 3424 - 3434, English, International magazine

  [Refereed]

  Scientific journal


• [Mannose trimming mechanism in endoplasmic reticulum-associated degradation of glycoproteins].

  Tetsuya Okada, Satoshi Ninagawa, Kazutoshi Mori

  Japanese Biochemical Society, Apr. 2016, Seikagaku. The Journal of Japanese Biochemical Society, 88(2) (2), 257 - 60, Japanese, Domestic magazine

  [Refereed]

  Scientific journal


• みにれびゅう 糖タンパク質の小胞体関連分解におけるマンノーストリミング機構

  岡田 徹也, 蜷川 暁, 森 和俊

  2016, 生化学, 88(2) (2), 257 - 260, Japanese

  [Refereed]


• Trypsin Sensitivity Assay to Study the Folding Status of Proteins.

  Ninagawa, S, Mori, K

  Lead, 2016, Bio-protocol, 6(19) (19), e1953

  [Refereed]


• PNGase Sensitivity Assay to Study the Folding Status of Proteins.

  Ninagawa, S, Mori, K

  Lead, 2016, Bio-protocol, 6(19) (19), e1952

  [Refereed]


• Derlin2/3およびHerp1/2はSEL1L依存的な構造異常タンパク質分解経路に必要である

  杉本 岳大, 蜷川 暁, 山野 晋平, 石川 時郎, 岡田 徹也, 武田 俊一, 森 和俊

  (公社)日本生化学会, Dec. 2015, 日本生化学会大会・日本分子生物学会年会合同大会講演要旨集, 88回・38回, [4T17 - 09(3P0384)], Japanese


• 糖鎖非依存小胞体関連分解経路によるシビアな構造異常糖タンパク質の強制分解

  蜷川 暁, 岡田 徹也, 住友 嘉樹, 堀本 賢, 杉本 岳大, 石川 時郎, 武田 俊一, 山本 卓, 神谷 由紀子, 加藤 晃一, 森 和俊

  (公社)日本生化学会, Dec. 2015, 日本生化学会大会・日本分子生物学会年会合同大会講演要旨集, 88回・38回, [2P0296] - [2P0296], Japanese


• Forcible destruction of severely misfolded mammalian glycoproteins by the non-glycoprotein ERAD pathway.

  Satoshi Ninagawa, Tetsuya Okada, Yoshiki Sumitomo, Satoshi Horimoto, Takehiro Sugimoto, Tokiro Ishikawa, Shunichi Takeda, Takashi Yamamoto, Tadashi Suzuki, Yukiko Kamiya, Koichi Kato, Kazutoshi Mori

  Lead, Nov. 2015, The Journal of cell biology, 211(4) (4), 775 - 84, English, International magazine

  [Refereed]

  Scientific journal


• 糖鎖依存的構造異常タンパク質分解に必須な糖鎖刈り込み機構を解明

  蜷川 暁, 加藤 晃一, 森 和俊

  Lead, 公益社団法人 日本農芸化学会, 2015, 化学と生物, 53(9) (9), 571 - 573, Japanese

  [Refereed]


• EDEM2 initiates mammalian glycoprotein ERAD by catalyzing the first mannose trimming step.

  Satoshi Ninagawa, Tetsuya Okada, Yoshiki Sumitomo, Yukiko Kamiya, Koichi Kato, Satoshi Horimoto, Tokiro Ishikawa, Shunichi Takeda, Tetsushi Sakuma, Takashi Yamamoto, Kazutoshi Mori

  Lead, Aug. 2014, The Journal of cell biology, 206(3) (3), 347 - 56, English, International magazine

  [Refereed]

  Scientific journal


• The unfolded protein response transducer ATF6 represents a novel transmembrane-type endoplasmic reticulum-associated degradation substrate requiring both mannose trimming and SEL1L protein.

  Satoshi Horimoto, Satoshi Ninagawa, Tetsuya Okada, Hibiki Koba, Takehiro Sugimoto, Yukiko Kamiya, Koichi Kato, Shunichi Takeda, Kazutoshi Mori

  Lead, Nov. 2013, The Journal of biological chemistry, 288(44) (44), 31517 - 27, English, International magazine

  [Refereed]

  Scientific journal


• ERにおける蛋白質品質管理システムに関する逆遺伝学的解析(Reverse Genetic Analysis of Protein Quality Control System in the ER)

  蜷川 暁, 住友 嘉樹, 岡田 徹也, 武田 俊一, 森 和俊

  (公社)日本生化学会, Sep. 2011, 日本生化学会大会プログラム・講演要旨集, 84回, 3S11p - 4, English


• SEL1L is required for endoplasmic reticulum-associated degradation of misfolded luminal proteins but not transmembrane proteins in chicken DT40 cell line.

  Satoshi Ninagawa, Tetsuya Okada, Shunichi Takeda, Kazutoshi Mori

  Lead, 2011, Cell structure and function, 36(2) (2), 187 - 95, English, Domestic magazine

  [Refereed]

  Scientific journal


• トリDT40細胞のERADの解析 Sel1Lノックアウトの影響(Analysis of ERAD in chicken DT40 cells: effects of Sel1L knockout)

  蜷川 暁, 岡田 徹也, 武田 俊一, 森 和俊

  (公社)日本生化学会, Dec. 2010, 日本生化学会大会・日本分子生物学会年会合同大会講演要旨集, 83回・33回, 4T10 - 5, English

• 小胞体マンノシダーゼEDEM2-S-S-TXNDC11複合体形成メカニズムの解明

  大下修一郎, 山代萌, 橋井則貴, LIEW Chia Yen, LIN Yen Ting, NI Chi Kung, 矢木宏和, 蜷川暁, 蜷川暁

  2024, 日本分子生物学会年会プログラム・要旨集(Web), 47th


• 小胞体は糖鎖を介した極めて巧妙なタンパク質の恒常性維持機構を有する

  蜷川暁

  2024, 日本分子生物学会年会プログラム・要旨集(Web), 47th


• 小胞体タンパク質構造形成関連因子群がタンパク質の分泌に与える影響の網羅的解析

  笛木茜, 津曲和哉, 松尾將生, 八尋錦之助, 鈴木悠太, 鹿島誠, 今見考志, 蜷川暁, 蜷川暁

  2024, 日本分子生物学会年会プログラム・要旨集(Web), 47th


• Mechanism of ER-localized glycoprotein quality control by N-glycan dependent folding and degradation related factors

  松尾將生, 津曲和哉, 鹿島誠, 今見考志, 蜷川暁

  2024, 日本糖質学会年会要旨集, 43rd


• UGGTs prevent premature degradation of unstable and misfolded glycoproteins in the endoplasmic reticulum

  蜷川暁, 蜷川暁, 松尾將生, DENG Ying, 阿曽伸哉, 松下和俊, 笛木茜, 斎藤俊介, 今見考志, 木塚康彦, 佐久間哲史, 山本卓, 矢木宏和, 加藤晃一, 加藤晃一, 加藤晃一, 森和俊

  2023, 日本糖質学会年会要旨集, 42nd


• UGGTs affect not only glycoprotein folding but also degradation

  松尾將生, DENG Ying, 矢木宏和, 矢木宏和, 津曲和哉, 笛木茜, 今見考志, 加藤晃一, 加藤晃一, 加藤晃一, 森和俊, 蜷川暁, 蜷川暁

  2023, 日本分子生物学会年会プログラム・要旨集(Web), 46th


• Folding or Degradation, Molecular Mechanisms Determining the Fate of Glycoproteins in the Endoplasmic Reticulum

  蜷川暁, 松尾將生, DENG Ying, 森和俊

  2023, 日本分子生物学会年会プログラム・要旨集(Web), 46th


• Functional analysis of Golgi-localized mannosidase MAN1B1.

  大下修一郎, 山代萌, 矢木宏和, LIEW Chia-Yen, NI Chi-Kung, 加藤晃一, 蜷川暁

  2023, 日本分子生物学会年会プログラム・要旨集(Web), 46th


• 小胞体における構造不全糖タンパク質の代謝タイミング決定の分子機構

  蜷川暁, 森和俊

  2023, 日本生化学会大会(Web), 96th


• Elucidating mechanisms underlying the robustness of the newly synthesized protein load in the endoplasmic reticulum

  蜷川暁, 森和俊

  2022, 日本分子生物学会年会プログラム・要旨集(Web), 45th


• タンパク質個性の基盤構造形成の場としての視座からの小胞体の機能解明

  蜷川暁, 森和俊

  2022, 日本生化学会大会(Web), 95th


• 小胞体におけるN型糖鎖による糖タンパク質の品質管理機構

  蜷川 暁

  Lead, 2021, Trends in glycoscience and glycotechnology, 33(191-193) (191-193), J55 - 62, Japanese


• N-glycan Dependent Protein Quality Control System in the Endoplasmic Reticulum

  蜷川暁

  Lead, 2021, Trends in Glycoscience and Glycotechnology (Web), 33(193) (193), E55 - E62


• Molecular mechanism of protein folding and degradation machinery in the mammalian endoplasmic reticulum

  蜷川暁

  Lead, 2021, 生化学, 93(4) (4)


• 小胞体関連分解因子EDEM2は,TXNDC11とジスルフィド結合を介した複合体を形成し,マンノシダーゼ活性を発揮する

  蜷川暁, GEORGE Ginto, 矢木宏和, 住友嘉樹, 石川時郎, 佐久間哲史, 山本卓, 今見考志, 石濱泰, 加藤晃一, 岡田徹也, 森和俊

  (公社)日本生化学会, 2020, 日本生化学会大会(Web), 93rd, [2Z01 - 027)], Japanese

• Folding or Degradation, Molecular Mechanisms Determining the Fate of Glycoproteins in the Endoplasmic Reticulum

  蜷川暁, 松尾將生, DENG Ying, 森和俊

  日本分子生物学会年会プログラム・要旨集(Web), 2023

  [Invited]


• Functional analysis of Golgi-localized mannosidase MAN1B1.

  大下修一郎, 山代萌, 矢木宏和, 矢木宏和, LIEW Chia-Yen, NI Chi-Kung, 加藤晃一, 蜷川暁

  日本分子生物学会年会プログラム・要旨集(Web), 2023


• 小胞体における構造不全糖タンパク質の代謝タイミング決定の分子機構

  蜷川暁, 森和俊

  日本生化学会大会(Web), 2023

  [Invited]


• UGGTs prevent premature degradation of unstable and misfolded glycoproteins in the endoplasmic reticulum

  蜷川暁, 松尾將生, DENG Ying, 阿曽伸哉, 松下和俊, 笛木茜, 斎藤俊介, 今見考志, 木塚康彦, 佐久間哲史, 山本卓, 矢木宏和, 加藤晃一, 森和俊

  日本糖質学会年会要旨集, 2023

  [Invited]


• Effects of the endoplasmic reticulum folding sensor UGGT on secretion

  笛木茜, 津曲和哉, 松尾將生, 今見考志, 蜷川暁

  日本分子生物学会年会プログラム・要旨集(Web), 2023


• UGGTs affect not only glycoprotein folding but also degradation

  松尾將生, DENG Ying, 矢木宏和, 矢木宏和, 津曲和哉, 笛木茜, 今見考志, 加藤晃一, 加藤晃一, 森和俊, 蜷川暁

  日本分子生物学会年会プログラム・要旨集(Web), 2023


• 平成から令和時代への小胞体「糖」タンパク質品質管理機構

  蜷川暁

  日本糖質学会年会要旨集, 2022

  [Invited]


• Elucidating mechanisms underlying the robustness of the newly synthesized protein load in the endoplasmic reticulum

  蜷川暁, 森和俊

  日本分子生物学会年会プログラム・要旨集(Web), 2022

  [Invited]


• タンパク質個性の基盤構造形成の場としての視座からの小胞体の機能解明

  蜷川暁, 森和俊

  日本生化学会大会(Web), 2022

  [Invited]


• The second-generation antipsychotic olanzapine induces aberrant intermolecular disulfide bonds of proinsulin

  蜷川暁, 多田誠一郎, 奥村正樹, 井ノ口健太, 木下岬, 金村進吾, 金村進吾, 今見考志, 梅澤元, 石川時郎, MACKIN Robert, 鳥居征司, 石濱泰, 稲葉謙次, 穴澤貴行, 長嶺敬彦, 森和俊

  日本細胞生物学会大会(Web), 2021


• The second generation antipsychotic olanzapine induces atypical development of diabetes due to aberrant disulfide bond formations of proinsulin in pancreatic beta cells

  蜷川暁, 多田誠一郎, 奥村正樹, 井ノ口健太, 木下岬, 金村進吾, 今見考志, 梅澤元, 石川時郎, MACKIN Robert, 鳥居征司, 石濱泰, 稲葉謙次, 穴澤貴行, 長嶺敬彦, 森和俊

  日本薬学会年会要旨集(Web), 2021


• The regulation by OS9 and XTP3B in ER-associated degradation depends on the degree of structural abnormality of degradation substrates

  松下和智, 蜷川暁, 杉本岳大, 岡田徹也, 石川時郎, 佐久間哲史, 山本卓, 森和俊

  日本糖質学会年会要旨集, 2021


• A mannosidase in the endoplasmic reticulum exerts its function by protein-protein association

  蜷川暁, GEORGE Ginto, 矢木宏和, 古川潤一, 橋井則貴, 石井明子, DENG Ying, 石川時郎, 今見考志, 石濱泰, 加藤晃一, 加藤晃一, 岡田徹也, 森和俊

  日本分子生物学会年会プログラム・要旨集(Web), 2021

  [Invited]


• Elucidation of molecular mechanisms and functions of N-glycan mannose trimming required for glycoprotein ER-associated degradation pathway

  蜷川暁, GEORGE Ginto, 矢木宏和, 住友嘉樹, 真木勇太, 石川時郎, 佐久間哲史, 山本卓, 今見考志, 石濱泰, 梶原康宏, 加藤晃一, 加藤晃一, 岡田徹也, 森和俊

  日本糖質学会年会要旨集, 2021

  [Invited]


• The second generation antipsychotic olanzapine induces ER-associated degradation of proinsulin due to its abberrant disulfide bond formation

  蜷川暁, 多田誠一郎, 井ノ口健太, 今見考志, 梅澤元, 石川時郎, 岡田徹也, MACKIN Robert, 鳥居征司, 石濱泰, 穴澤貴行, 長嶺敬彦, 森和俊

  日本薬学会年会要旨集(CD-ROM), Mar. 2020, Japanese, (公社)日本薬学会


• 遺伝子破壊法を用いた小胞体関連分解機構の解明

  蜷川暁

  日本生化学会大会(Web), 2020

  [Invited]


• 小胞体関連分解因子EDEM2は,TXNDC11とジスルフィド結合を介した複合体を形成し,マンノシダーゼ活性を発揮する

  蜷川暁, GEORGE Ginto, 矢木宏和, 住友嘉樹, 石川時郎, 佐久間哲史, 山本卓, 今見考志, 石濱泰, 加藤晃一, 加藤晃一, 岡田徹也, 森和俊

  日本生化学会大会(Web), 2020

  [Invited]


• An enzymatic activity of EDEM2 is endowed by TXNDC11 connected via a disulfide bond

  蜷川暁, GEORGE Ginto, 矢木宏和, 斎藤泰輝, 住友嘉樹, 石川時郎, 佐久間哲史, 山本卓, 今見考志, 石濱泰, 加藤晃一, 加藤晃一, 岡田徹也, 森和俊

  日本糖質学会年会要旨集, 2020


• メダカを用いた小胞体ストレスに起因するアポトーシス分子機構の解析

  陳 炳碩, 石川 時郎, 蜷川 暁, 岡田 徹也, 森 和俊

  日本生化学会大会プログラム・講演要旨集, Sep. 2019, Japanese, (公社)日本生化学会


• タンパク質の運命を制御する生体システムと疾病のフロンティア 第二世代抗精神病薬オランザピンは、副作用としてProinsulinの適切なジスルフィド結合形成を阻害し糖尿病を誘発する

  蜷川 暁, 岡田 徹也, 梅澤 元, 石川 時郎, 鳥居 征司, Mackin Robert, 今見 考志, 石濱 泰, 長嶺 敬彦, 森 和俊

  日本生化学会大会プログラム・講演要旨集, Sep. 2019, Japanese, (公社)日本生化学会


• 小胞体ストレスセンサーATF6αはジスルフィド結合を介して2種類の二量体を形成する(Endoplasmic reticulum stress sensor ATF6α forms two types of dimers via disulfide-bonding)

  古場 玲, 金 聖宇, 岡田 徹也, 石川 時郎, 蜷川 暁, 森 和俊

  日本生化学会大会プログラム・講演要旨集, Sep. 2019, Japanese, (公社)日本生化学会


• 小胞体膜タンパク質Seipinの神経変性誘導変異体に起因する小胞体ストレス発生機構の解析

  齊藤 峻介, 石川 時郎, 蜷川 暁, 岡田 徹也, 森 和俊

  日本生化学会大会プログラム・講演要旨集, Sep. 2019, Japanese, (公社)日本生化学会


• N型糖鎖のマンノーストリミングを基軸とした小胞体関連分解の解析

  蜷川暁

  細胞内糖修飾の統合的ケミカルバイオロジー講演要旨集 理研シンポジウム 平成31年, 2019

  [Invited]


• 高等動物におけるN型糖鎖トリミングを基軸とした小胞体関連分解機構の解析

  蜷川暁, GEORGE Ginto, 矢木宏和, 住友嘉樹, 神谷由紀子, 石川時郎, 武田俊一, 佐久間哲史, 山本卓, 加藤晃一, 加藤晃一, 岡田徹也, 森和俊

  日本糖質学会年会要旨集, 2019

  [Invited]


• 小胞体タンパク質品質管理機構の中心を担うN型糖鎖依存/非依存小胞体関連分解経路の分子メカニズムの解析

  蜷川暁, GINTO George, 矢木宏和, 今見考志, 石濱泰, 石川時郎, 佐久間哲史, 山本卓, 武田俊一, 加藤晃一, 加藤晃一, 岡田徹也, 森和俊

  日本分子生物学会年会プログラム・要旨集(Web), 2019

  [Invited]


• 第二世代抗精神病薬オランザピンがインスリン分泌不全を引き起こし、糖尿病を誘起する分子メカニズムの解析

  蜷川 暁, 岡田 徹也, 今見 考志, 梅澤 元, 石川 時郎, 鳥居 征司, Mackin Robert, 石濱 泰, 長嶺 敬彦, 森 和俊

  日本生化学会大会プログラム・講演要旨集, Sep. 2018, Japanese, (公社)日本生化学会


• 第二世代抗精神病薬オランザピンが、副作用として糖尿病を引き起こす分子メカニズムの解析 インスリンは、適切なジスルフィド結合形成を阻害され、小胞体関連分解により処理される

  蜷川 暁, 岡田 徹也, 梅澤 元, 石川 時郎, 鳥居 征司, Mackin Robert, 長嶺 敬彦, 森 和俊

  生命科学系学会合同年次大会, Dec. 2017, Japanese, 生命科学系学会合同年次大会運営事務局


• α-ジストログリカンのラミニン結合性糖鎖形成するポストリン酸修飾の構造解析

  矢木 宏和, Kuo Chu-Wei, 尾林 卓幸, 蜷川 暁, Khoo Kay-Hooi, 加藤 晃一

  日本薬学会年会要旨集, Mar. 2017, Japanese, (公社)日本薬学会


• オルガネラ局域の間取り図 分解執行局域における新展開

  蜷川 暁, 岡田 徹也, 住友 嘉樹, 堀本 賢, 鈴木 匡, 武田 俊一, 佐久間 哲史, 山本 卓, 神谷 由紀子, 加藤 晃一, 森 和俊

  日本細胞生物学会大会講演要旨集, May 2016, Japanese, (一社)日本細胞生物学会


• 糖鎖非依存小胞体関連分解経路によるシビアな構造異常糖タンパク質の強制分解

  蜷川 暁, 岡田 徹也, 住友 嘉樹, 堀本 賢, 杉本 岳大, 石川 時郎, 武田 俊一, 山本 卓, 神谷 由紀子, 加藤 晃一, 森 和俊

  日本生化学会大会・日本分子生物学会年会合同大会講演要旨集, Dec. 2015, Japanese, (公社)日本生化学会


• Derlin2/3およびHerp1/2はSEL1L依存的な構造異常タンパク質分解経路に必要である

  杉本 岳大, 蜷川 暁, 山野 晋平, 石川 時郎, 岡田 徹也, 武田 俊一, 森 和俊

  日本生化学会大会・日本分子生物学会年会合同大会講演要旨集, Dec. 2015, Japanese, (公社)日本生化学会


• EDEM1/2/3 are alpha 1,2-mannosidases essential for endoplasmic reticulum-associated degradation of glycoproteins

  Satoshi Ninagawa, Tetsuya Okada, Yoshiki Sumitomo, Yukiko Kamiya, Satoshi Horimoto, Tokiro Ishikawa, Shunichi Takeda, Tetsushi Sakuma, Takashi Yamamoto, Koichi Kato, Kazutoshi Mori

  GLYCOBIOLOGY, Nov. 2014, English


• ERにおける蛋白質品質管理システムに関する逆遺伝学的解析(Reverse Genetic Analysis of Protein Quality Control System in the ER)

  蜷川 暁, 住友 嘉樹, 岡田 徹也, 武田 俊一, 森 和俊

  日本生化学会大会プログラム・講演要旨集, Sep. 2011, English, (公社)日本生化学会

  [Invited]


• トリDT40細胞のERADの解析 Sel1Lノックアウトの影響(Analysis of ERAD in chicken DT40 cells: effects of Sel1L knockout)

  蜷川 暁, 岡田 徹也, 武田 俊一, 森 和俊

  日本生化学会大会・日本分子生物学会年会合同大会講演要旨集, Dec. 2010, English, (公社)日本生化学会


• 小胞体関連分解構成因子Derlinsに結合するタンパク質の同定と解析

  NINAGAWA SATOSHI, OKADA TETSUYA, ODA YUKAKO, OKAWA KATSUYA, MORI KAZUTOSHI

  生化学, 2007, Japanese

• 日本糖質学会

  Apr. 2014 - Present


• 日本細胞生物学会

  Apr. 2010 - Present


• 日本分子生物学会

  Apr. 2010 - Present


• 日本生化学会

  Apr. 2008 - Present

• Interleukin-18の細胞外発現とその機能制御

  蜷川 暁

  公益財団法人 持田記念医学薬学振興財団, Dec. 2024 - Dec. 2025


• 哺乳動物の小胞体関連分解における小胞体内イベントの全容解明

  森 和俊, 蜷川 暁

  日本学術振興会, 科学研究費助成事業, 基盤研究(A), 京都大学, Apr. 2022 - Mar. 2025


• αジストログリカノパチー発症の新規分子基盤の解明

  蜷川 暁

  公益財団法人 内藤記念科学振興財団, Dec. 2022 - Sep. 2024


• α-Dystroglycanの機能的O型糖鎖形成に対するN型糖鎖のマンノース切除酵素の役割の解明

  蜷川 暁

  公益財団法人 小林財団, 第9回研究助成, Mar. 2021 - May 2024


• 小胞体局在性糖タンパク質品質管理機構の確立

  蜷川 暁

  公益財団法人 長瀬研究振興財団, Apr. 2023 - Mar. 2024


• N型糖鎖依存小胞体関連分解における小胞体内イベントの全容解明

  蜷川 暁

  公益財団法人 武田科学振興財団, ライフサイエンス研究助成, Oct. 2020 - May 2023


• Analysis of sorting zones involved in protein quality control in the endoplasmic reticulum

  Mori Kazutoshi

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research, Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area), Kyoto University, Jun. 2017 - Mar. 2022

  In this study, the following results were obtained for three organelle zones. Sorting zone: we visualized the intracellular transport of the endoplasmic reticulum (ER) stress sensors ATF6 and IRE1 and identified their regulators. Degradation zone: our analysis revealed that Derlin1 and Derlin2 can form different complexes that play different roles in protein degradation in the ER. We also found that Derlin1 and Derlin2 are essential for neurite outgrowth in the central nervous system and that TXNDC11 is essential for mannose trimming in glycoprotein degradation. Mitochondria-ER communication zone: we found that an ER-resident protein PERK is indispensable for mitochondrial biogenesis during brown adipocyte differentiation.


• Analysis of how misfolded proteins are delivered to retrotranslocation complex for ER-associated degradation

  Ninagawa Satoshi

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research, Grant-in-Aid for Scientific Research (C), Kyoto University, 01 Apr. 2018 - 31 Mar. 2021

  In the endoplasmic reticulum (ER) secretory and membrane proteins are synthesized. During the synthesis, some proteins cannot fold properly. Such misfolded proteins are degraded via proteasome in the cytosol. In this study, I focus on how misfolded proteins are delivered from ER lumen to ER membrane and revealed the redundant roles of OS9 and XTP3B for substrate degradation.


• 群馬大学, 生体調節研究所「内分泌・代謝学共同研究拠点」共同研究, Apr. 2018 - Mar. 2019, Principal investigator


• Insight into how severely misfolded proteins are degraded in the ER

  Ninagawa Satoshi

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Young Scientists (B), Grant-in-Aid for Young Scientists (B), Kyoto University, 01 Apr. 2016 - 31 Mar. 2018

  ER-associated degradation (ERAD) is composed of glycoprotein and non-glycoprotein degradation pathways. By inhibition of glycoprotein degradation pathway, native but unstable or somewhat unfolded glycoproteins were stabilized, but degradation of severely misfolded glycoproteins was delayed only at early chase periods, but they were eventually degraded as in wild-type cells. We analyzed these molecular mechanisms. We tried to knocked out the candidate gene, which target substrates from glycoprotein degradation pathway to non-glycoprotein degradation pathway, and found that the gene is essential for cells, indicating its importance. Using PITCh method, its low expression strains were established. The mutant strain includes some mutations in genomic level. We found that the strain is defective to degrade severely misfolded non-glycoproteins.


• Elucidation of regulation mechanism of the endoplasmic reticulum membrane-bound transcription factor ATF6 and its role in the evolution of vertebrates

  Mori Kazutoshi, OKADA Tetsuya, ISHIKAWA Tokiro, TAKEDA Shunichi, YAMAMOTO Takashi, KATO Koichi, NINAGAWA Satoshi

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research, Grant-in-Aid for Scientific Research (B), Kyoto University, Apr. 2014 - Mar. 2017

  The endoplasmic reticulum (ER) membrane-bound transcription factor ATF6 is a protein with a short half-life of 2 h and constitutively subjected to glycan-dependent ER-associated degradation, although it functions as an ER stress sensor and transducer. I analyzed mannose trimming enzymes which are important for this degradation mechanism and discovered that the trimming of M9 to M8 is carried out by EDEM2 and that the trimming of M8 to M7 is carried out by EDEM1/3 (EDEM3 plays a major role). These results have completely renewed the previous model.


• Analysis of ER-associated degradation components by multiple knockout cell lines using TALEN

  Ninagawa Satoshi

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Young Scientists (B), Grant-in-Aid for Young Scientists (B), 01 Apr. 2014 - 31 Mar. 2016

  Glycoproteins misfolded in the endoplasmic reticulum (ER) are subjected to ER-associated glycoprotein degradation. In this process, N-glycan mannose trimming is required. We clearly showed that this process is initiated by EDEM2 and completed by EDEM3/EDEM1. We also demonstrated that higher eukaryotes are able to extract severely misfolded glycoproteins from glycoprotein ERAD and target them to the non-glycoprotein ERAD pathway to maintain the homeostasis of the ER.


• 小胞体関連分解構成因子Derlinsに結合するタンパク質の同定と解析

  蜷川 暁

  日本学術振興会, 科学研究費助成事業 特別研究員奨励費, 特別研究員奨励費, 京都大学, 2008 - 2009

  小胞体関連分解とは小胞体において正しく立体構造をとることができないタンパク質を小胞体から細胞質に逆行輸送した後、細胞質のプロテアソームで分解する機構である。 研究計画に沿って研究を進めていたが、RNAi法を用いた遺伝子抑制実験では、きちんとした実験結果が出にくいということが判明したため、遺伝子破壊法であるknock out実験によって研究を進めた。具体的には、組み換えが非常におこりやすく遺伝子破壊しやすい、ニワトリBリンパ球細胞を用いて解析をした。現在までに小胞体関連分解の中心因子であるSellLや他3種のKnock out細胞の作成に成功した。 SellL-/-細胞は、小胞体関連分解の基質であるNHK(NNN)の分解遅延が見られた。また予想された通り糖鎖を持たないNHK(QQQ)の分解遅延も確認されている。これらのことによりDT40を用いた遺伝子破壊は期待された効果を得ており、今後、簡単に遺伝子操作できることから様々な遺伝子のknock outを得、解析することができ、現在までにきちんと調べられていなかった遺伝子もどのように4小胞体関連分解に関わっているかきちんと調べることができる。コントロールはとれてきたので、今後機能未知の遺伝子に関しても詳細に調べて行きたい。

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