Research activity information

• Oct. 2018 International Congress on Pure and Applied Chemistry Langkawi 2018, Plenary Lecture Award（International Congress on Pure and Applied Chemistry Langkawi 2018）, Membrane proteins in action; Structural and functional dynamics revealed by time-resolved measurements.

  KIMURA Tetsunari

  International society


• Jun. 2015 Protein Science Society of Japan, 蛋白質科学会若手奨励賞, マイクロ流体フローを用いた一酸化窒素還元酵素の触媒反応の動的分光観察

  KIMURA TETSUNARI

  Japan society

• Oxygen-evolving photosystem II structures during S1-S2-S3 transitions.

  Hongjie Li, Yoshiki Nakajima, Eriko Nango, Shigeki Owada, Daichi Yamada, Kana Hashimoto, Fangjia Luo, Rie Tanaka, Fusamichi Akita, Koji Kato, Jungmin Kang, Yasunori Saitoh, Shunpei Kishi, Huaxin Yu, Naoki Matsubara, Hajime Fujii, Michihiro Sugahara, Mamoru Suzuki, Tetsuya Masuda, Tetsunari Kimura, Tran Nguyen Thao, Shinichiro Yonekura, Long-Jiang Yu, Takehiko Tosha, Kensuke Tono, Yasumasa Joti, Takaki Hatsui, Makina Yabashi, Minoru Kubo, So Iwata, Hiroshi Isobe, Kizashi Yamaguchi, Michihiro Suga, Jian-Ren Shen

  Photosystem II (PSII) catalyses the oxidation of water through a four-step cycle of Si states (i = 0-4) at the Mn4CaO5 cluster1-3, during which an extra oxygen (O6) is incorporated at the S3 state to form a possible dioxygen4-7. Structural changes of the metal cluster and its environment during the S-state transitions have been studied on the microsecond timescale. Here we use pump-probe serial femtosecond crystallography to reveal the structural dynamics of PSII from nanoseconds to milliseconds after illumination with one flash (1F) or two flashes (2F). YZ, a tyrosine residue that connects the reaction centre P680 and the Mn4CaO5 cluster, showed structural changes on a nanosecond timescale, as did its surrounding amino acid residues and water molecules, reflecting the fast transfer of electrons and protons after flash illumination. Notably, one water molecule emerged in the vicinity of Glu189 of the D1 subunit of PSII (D1-E189), and was bound to the Ca2+ ion on a sub-microsecond timescale after 2F illumination. This water molecule disappeared later with the concomitant increase of O6, suggesting that it is the origin of O6. We also observed concerted movements of water molecules in the O1, O4 and Cl-1 channels and their surrounding amino acid residues to complete the sequence of electron transfer, proton release and substrate water delivery. These results provide crucial insights into the structural dynamics of PSII during S-state transitions as well as O-O bond formation.

  Feb. 2024, Nature, 626(7999) (7999), 670 - 677, English, International magazine

  Scientific journal


• Trapping of a Mononitrosyl Nonheme Intermediate of Nitric Oxide Reductase by Cryo-Photolysis of Caged Nitric Oxide.

  Hanae Takeda, Kanji Shimba, Masaki Horitani, Tetsunari Kimura, Takashi Nomura, Minoru Kubo, Yoshitsugu Shiro, Takehiko Tosha

  Characterization of short-lived reaction intermediates is essential for elucidating the mechanism of the reaction catalyzed by metalloenzymes. Here, we demonstrated that the photolysis of a caged compound under cryogenic temperature followed by thermal annealing is an invaluable technique for trapping of short-lived reaction intermediates of metalloenzymes through the study of membrane-integrated nitric oxide reductase (NOR) that catalyzes reductive coupling of two NO molecules to N2O at its heme/nonheme FeB binuclear center. Although NO produced by the photolysis of caged NO did not react with NOR under cryogenic temperature, annealing to ∼160 K allowed NO to diffuse and react with NOR, which was evident from the appearance of EPR signals assignable to the S = 3/2 state. This indicates that the nonheme FeB-NO species can be trapped as the intermediate. Time-resolved IR spectroscopy with the use of the photolysis of caged NO as a reaction trigger showed that the intermediate formed at 10 μs gave the NO stretching frequency at 1683 cm-1 typical of nonheme Fe-NO, confirming that the combination of the cryo-photolysis of caged NO and annealing enabled us to trap the reaction intermediate. Thus, the cryo-photolysis of the caged compound has great potential for the characterization of short-lived reaction intermediates.

  Feb. 2023, The journal of physical chemistry. B, 127(4) (4), 846 - 854, English, International magazine

  Scientific journal


• Short-lived intermediate in N2O generation by P450 NO reductase captured by time-resolved IR spectroscopy and XFEL crystallography

  Takashi Nomura, Tetsunari Kimura, Yusuke Kanematsu, Daichi Yamada, Keitaro Yamashita, Kunio Hirata, Go Ueno, Hironori Murakami, Tamao Hisano, Raika Yamagiwa, Hanae Takeda, Chai Gopalasingam, Ryota Kousaka, Sachiko Yanagisawa, Osami Shoji, Takashi Kumasaka, Masaki Yamamoto, Yu Takano, Hiroshi Sugimoto, Takehiko Tosha, Minoru Kubo, Yoshitsugu Shiro

  Nitric oxide (NO) reductase from the fungus Fusarium oxysporum is a P450-type enzyme (P450nor) that catalyzes the reduction of NO to nitrous oxide (N₂O) in the global nitrogen cycle. In this enzymatic reaction, the heme-bound NO is activated by the direct hydride transfer from NADH to generate a short-lived intermediate (I), a key state to promote N–N bond formation and N–O bond cleavage. This study applied time-resolved (TR) techniques in conjunction with photolabile-caged NO to gain direct experimental results for the characterization of the coordination and electronic structures of I. TR freeze-trap crystallography using an X-ray free electron laser (XFEL) reveals highly bent Fe–NO coordination in I, with an elongated Fe–NO bond length (Fe–NO = 1.91 Å, Fe–N–O = 138°) in the absence of NAD⁺. TR-infrared (IR) spectroscopy detects the formation of I with an N–O stretching frequency of 1,290 cm⁻¹ upon hydride transfer from NADH to the Fe³⁺–NO enzyme via the dissociation of NAD⁺ from a transient state, with an N–O stretching of 1,330 cm⁻¹ and a lifetime of ca. 16 ms. Quantum mechanics/molecular mechanics calculations, based on these crystallographic and IR spectroscopic results, demonstrate that the electronic structure of I is characterized by a singly protonated Fe³⁺–NHO^•− radical. The current findings provide conclusive evidence for the N₂O generation mechanism via a radical–radical coupling of the heme nitroxyl complex with the second NO molecule.

  Proceedings of the National Academy of Sciences, May 2021, Proceedings of the National Academy of Sciences, 118(21) (21), e2101481118 - e2101481118

  Scientific journal


• Capturing structural changes of the S1 to S2 transition of photosystem II using time-resolved serial femtosecond crystallography.

  Hongjie Li, Yoshiki Nakajima, Takashi Nomura, Michihiro Sugahara, Shinichiro Yonekura, Siu Kit Chan, Takanori Nakane, Takahiro Yamane, Yasufumi Umena, Mamoru Suzuki, Tetsuya Masuda, Taiki Motomura, Hisashi Naitow, Yoshinori Matsuura, Tetsunari Kimura, Kensuke Tono, Shigeki Owada, Yasumasa Joti, Rie Tanaka, Eriko Nango, Fusamichi Akita, Minoru Kubo, So Iwata, Jian-Ren Shen, Michihiro Suga

  Photosystem II (PSII) catalyzes light-induced water oxidation through an S i -state cycle, leading to the generation of di-oxygen, protons and electrons. Pump-probe time-resolved serial femtosecond crystallography (TR-SFX) has been used to capture structural dynamics of light-sensitive proteins. In this approach, it is crucial to avoid light contamination in the samples when analyzing a particular reaction intermediate. Here, a method for determining a condition that avoids light contamination of the PSII microcrystals while minimizing sample consumption in TR-SFX is described. By swapping the pump and probe pulses with a very short delay between them, the structural changes that occur during the S1-to-S2 transition were examined and a boundary of the excitation region was accurately determined. With the sample flow rate and concomitant illumination conditions determined, the S2-state structure of PSII could be analyzed at room temperature, revealing the structural changes that occur during the S1-to-S2 transition at ambient temperature. Though the structure of the manganese cluster was similar to previous studies, the behaviors of the water molecules in the two channels (O1 and O4 channels) were found to be different. By comparing with the previous studies performed at low temperature or with a different delay time, the possible channels for water inlet and structural changes important for the water-splitting reaction were revealed.

  May 2021, IUCrJ, 8(Pt 3) (Pt 3), 431 - 443, English, International magazine

  Scientific journal


• Time-resolved serial femtosecond crystallography reveals early structural changes in channelrhodopsin.

  Kazumasa Oda, Takashi Nomura, Takanori Nakane, Keitaro Yamashita, Keiichi Inoue, Shota Ito, Johannes Vierock, Kunio Hirata, Andrés D Maturana, Kota Katayama, Tatsuya Ikuta, Itsuki Ishigami, Tamaki Izume, Rie Umeda, Ryuun Eguma, Satomi Oishi, Go Kasuya, Takafumi Kato, Tsukasa Kusakizako, Wataru Shihoya, Hiroto Shimada, Tomoyuki Takatsuji, Mizuki Takemoto, Reiya Taniguchi, Atsuhiro Tomita, Ryoki Nakamura, Masahiro Fukuda, Hirotake Miyauchi, Yongchan Lee, Eriko Nango, Rie Tanaka, Tomoyuki Tanaka, Michihiro Sugahara, Tetsunari Kimura, Tatsuro Shimamura, Takaaki Fujiwara, Yasuaki Yamanaka, Shigeki Owada, Yasumasa Joti, Kensuke Tono, Ryuichiro Ishitani, Shigehiko Hayashi, Hideki Kandori, Peter Hegemann, So Iwata, Minoru Kubo, Tomohiro Nishizawa, Osamu Nureki

  Channelrhodopsins (ChRs) are microbial light-gated ion channels utilized in optogenetics to control neural activity with light . Light absorption causes retinal chromophore isomerization and subsequent protein conformational changes visualized as optically distinguished intermediates, coupled with channel opening and closing. However, the detailed molecular events underlying channel gating remain unknown. We performed time-resolved serial femtosecond crystallographic analyses of ChR by using an X-ray free electron laser, which revealed conformational changes following photoactivation. The isomerized retinal adopts a twisted conformation and shifts toward the putative internal proton donor residues, consequently inducing an outward shift of TM3, as well as a local deformation in TM7. These early conformational changes in the pore-forming helices should be the triggers that lead to opening of the ion conducting pore.

  Mar. 2021, eLife, 10, English, International magazine

  Scientific journal


• Role of Zinc Oxide Nanoparticles Synthesized by Fenugreek Seeds Extract as Anticancer Agent: In Vitro and In Vivo Studies.

  Aliaa M Radwan, Eman F Aboelfetoh, Tetsunari Kimura, Tarek M Mohamed, Mai M El-Keiy

  BACKGROUND: Zinc oxide nanoparticles (ZnO NPs) are one of the metal oxide nanoparticles, which have attracted the interest of the researchers due to their biocompatibility, easily surface functionalization, and cancer targeting. OBJECTIVE: This study was designated to investigate the potential antitumor activity of the biologically synthesized ZnO NPs alone or in combination with doxorubicin using Ehrlich ascites carcinoma (EAC) model. METHODS: In this study, ZnO NPs were prepared by green approach using fenugreek seeds extract as reducing and capping agent then characterized by scanning electron microscope (SEM), energy dispersive x-ray (EDX), X-ray diffraction (XRD), UV-V spectroscopy, and transmission electron microscope (TEM). The prepared nanoparticles were tested for in vitro and in vivo studies using different parameters. RESULTS: Zinc oxide nanoparticles were determined to have cytotoxicity against different cancer cell lines with lower toxicity against normal one. Moreover, the in vivo study, demonstrated that the intraperitoneal injection of ZnO NPs alone or combined with doxorubicin in EAC mice inhibited the proliferation and growth of EAC by decreasing the ascetic volume and viable tumor cell count. This anti-proliferative efficiency of ZnO NPs was due to cell cycle arrest at G0/G1 phase and induction of apoptosis via upregulating the expression of caspase-3 and Bax and downregulating the expression of Bcl-2. CONCLUSION: Our findings indicated that the biologically synthesized ZnO NPs may be a promising nanomedicine therapy for cancer treatment in the future.

  Jan. 2021, Anti-cancer agents in medicinal chemistry, English, International magazine

  Scientific journal


• Functional Assembly of Caenorhabditis elegans Cytochrome b-2 (Cecytb-2) into Phospholipid Bilayer Nanodisc with Enhanced Iron Reductase Activity.

  Hamed A Abosharaf, Yuki Sakamoto, Aliaa M Radwan, Keisuke Yuzu, Mika Fujimura, Thoria Diab, Tarek M Mohamed, Eri Chatani, Tetsunari Kimura, Motonari Tsubaki

  Among seven homologs of cytochrome b561 in a model organism C. elegans, Cecytb-2 was confirmed to be expressed in digestive organs and was considered as a homolog of human Dcytb functioning as a ferric reductase. Cecytb-2 protein was expressed in Pichia pastoris cells, purified, and reconstituted into a phospholipid bilayer nanodisc. The reconstituted Cecytb-2 in nanodisc environments was extremely stable and more reducible with ascorbate than in a detergent-micelle state. We confirmed the ferric reductase activity of Cecytb-2 by analyzing the oxidation of ferrous heme upon addition of ferric substrate under anaerobic conditions, where clear and saturable dependencies on the substrate concentrations following the Michaelis-Menten equation were observed. Further, we confirmed that the ferric substrate was converted to a ferrous state by using a nitroso-PSAP assay. Importantly, we observed that the ferric reductase activity of Cecytb-2 became enhanced in the phospholipid bilayer nanodisc.

  Jan. 2021, Biomolecules, 11(1) (1), English, International magazine

  Scientific journal

  Link to Kobe Univ. Repository (Kernel)


• Timing of NO Binding and Protonation in the Catalytic Reaction of Bacterial Nitric Oxide Reductase as Established by Time-Resolved Spectroscopy

  Hanae Takeda, Tetsunari Kimura, Takashi Nomura, Masaki Horitani, Azusa Yokota, Akiko Matsubayashi, Shoko Ishii, Yoshitsugu Shiro, Minoru Kubo, Takehiko Tosha

  The Chemical Society of Japan, Jul. 2020, Bulletin of the Chemical Society of Japan, 93(7) (7), 825 - 833

  [Refereed]

  Scientific journal


• Direct measurements of ferric reductase activity of human 101F6 and its enhancement upon reconstitution into phospholipid bilayer nanodisc.

  Mohammed El Behery, Mika Fujimura, Tetsunari Kimura, Motonari Tsubaki

  We studied human 101F6 protein to clarify its physiological function as a ferric reductase and its relationship to tumor suppression activity. We found for the first time that purified 101F6 both in detergent micelle state and in phospholipid bilayer nanodisc state has an authentic ferric reductase activity by single turnover kinetic analyses. The kinetic analysis on the ferrous heme oxidation of reduced 101F6 upon the addition of a ferric substrate, ferric ammonium citrate (FAC), showed concentration-dependent accelerations of its reaction with reasonable values of KM and Vmax. We further verified the authenticity of the ferric reductase activity of 101F6 using nitroso-PSAP as a Fe2+-specific colorimetric chelator. 101F6 in nanodisc state showed higher efficiency for FAC than in detergent micelle state.

  Mar. 2020, Biochemistry and biophysics reports, 21, 100730 - 100730, English, International magazine

  [Refereed]

  Scientific journal

  Link to Kobe Univ. Repository (Kernel)


• An oxyl/oxo mechanism for oxygen-oxygen coupling in PSII revealed by an x-ray free-electron laser.

  Suga M, Akita F, Yamashita K, Nakajima Y, Ueno G, Li H, Yamane T, Hirata K, Umena Y, Yonekura S, Yu LJ, Murakami H, Nomura T, Kimura T, Kubo M, Baba S, Kumasaka T, Tono K, Yabashi M, Isobe H, Yamaguchi K, Yamamoto M, Ago H, Shen JR

  Oct. 2019, Science (New York, N.Y.), 366(6463) (6463), 334 - 338, English

  [Refereed]

  Scientific journal


• Elucidation of molecular functions of human tumor suppressor protein 101F6 by reconstitution into phospholipid bilayer nanodiscs

  El Behery, Mohammed, Asada, Akikazu, Takeuchi, Fusako, Kimura, Tetsunari, Chatani, Eri, TSUBAKI MOTONARI

  Chemical Society of the Philippines, Jun. 2019, KIMIKA Journal of the Kapisanang Kimika ng Pilipinas, 30(1) (1), 1 - 3, English

  [Refereed]

  Scientific journal

  Link to Kobe Univ. Repository (Kernel)


• Vibrational and Molecular Properties of Mg2+ Binding and Ion Selectivity in the Magnesium Channel MgtE

  Kimura, T, Lorenz-Fonfria, V.A, Douki, S, Motoki, H, Ishitani, R, Nureki, O, Higashi, M, Furutani, Y

  Magnesium ions (Mg2+) are crucial for various biological processes. A bacterial Mg2+ channel, MgtE, tightly regulates the intracellular Mg2+ concentration. Previous X-ray crystal structures showed that MgtE forms a dimeric structure composed of a total of 10 transmembrane α helices forming a central pore, and intracellular soluble domains constituting a Mg2+ sensor. The ion selectivity for Mg2+ over Ca2+ resides at a central cavity in the transmembrane pore of MgtE, involving a conserved aspartate residue (Asp432) from each monomer. Here, we applied ion-exchange-induced difference FTIR spectroscopy to analyze the interactions between MgtE and divalent cations, Mg2+ and Ca2+. Using site-directed mutagenesis, vibrational bands at 1421 (Mg2+), 1407 (Mg2+), ∼1440 (Ca2+), and 1390 (Ca2+) cm-1 were assigned to symmetric carboxylate stretching modes of Asp432, involved in the ion coordination. Conservative modifications of the central cavity by Asp432Glu or Ala417Leu mutations resulted in the disappearance of the Mg2+-sensitive carboxylate bands, suggesting a highly optimized geometry for accommodating a Mg2+ ion. The dependency of the vibrational changes on Mg2+ and Ca2+ concentrations revealed the presence of a two different classes of binding sites: a high affinity site for Mg2+ ( Kd ≈ 0.3 mM) with low Ca2+ affinity ( Kd ≈ 80 mM), and a medium affinity site for Mg2+ ( Kd ≈ 2 mM) and Ca2+ ( Kd ≈ 6 mM), tentatively assigned to the central cavity and the sensor domain, respectively. With the aid of molecular dynamics simulation and normal-mode analysis by quantum chemistry, we confirm that changes in carboxylate bands of the high affinity binding site originate from Asp432 in the central cavity.

  Oct. 2018, J. Phys. Chem. B, 122(42) (42), 9681 - 9696, English, International magazine

  [Refereed]

  Scientific journal


• Vibrational and molecular properties of Mg2+ binding and selectivity in the magnesium channel MgtE

  Tetsunari KIMURA, Victor A, LORENZ-FONFRIA, Shintaro DOUKI, Hideyoshi MOTOKI, Ryuichiro ISHITANI, Osamu NUREKI, Masahiro HIGASHI, Yuji FURUTANI

  Magnesium ions (Mg2+) are crucial for various biological processes. A bacterial Mg2+ channel, MgtE, tightly regulates the intracellular Mg2+ concentration. Previous X-ray crystal structures showed that MgtE forms a dimeric structure composed of a total of 10 transmembrane α helices forming a central pore, and intracellular soluble domains constituting a Mg2+ sensor. The ion selectivity for Mg2+ over Ca2+ resides at a central cavity in the transmembrane pore of MgtE, involving a conserved aspartate residue (Asp432) from each monomer. Here, we applied ion-exchange-induced difference FTIR spectroscopy to analyze the interactions between MgtE and divalent cations, Mg2+ and Ca2+. Using site-directed mutagenesis, vibrational bands at 1421 (Mg2+), 1407 (Mg2+), ∼1440 (Ca2+), and 1390 (Ca2+) cm-1 were assigned to symmetric carboxylate stretching modes of Asp432, involved in the ion coordination. Conservative modifications of the central cavity by Asp432Glu or Ala417Leu mutations resulted in the disappearance of the Mg2+-sensitive carboxylate bands, suggesting a highly optimized geometry for accommodating a Mg2+ ion. The dependency of the vibrational changes on Mg2+ and Ca2+ concentrations revealed the presence of a two different classes of binding sites: a high affinity site for Mg2+ ( Kd ≈ 0.3 mM) with low Ca2+ affinity ( Kd ≈ 80 mM), and a medium affinity site for Mg2+ ( Kd ≈ 2 mM) and Ca2+ ( Kd ≈ 6 mM), tentatively assigned to the central cavity and the sensor domain, respectively. With the aid of molecular dynamics simulation and normal-mode analysis by quantum chemistry, we confirm that changes in carboxylate bands of the high affinity binding site originate from Asp432 in the central cavity.

  2018, J. Phys. Chem. B., 122(42) (42), 9681 - 9696, English, International magazine

  [Refereed]

  Scientific journal


• Ultraprecision cutting of single-crystal calcium fluoride for fabricating micro flow cells

  Ryo Komiya, Tetsunari Kimura, Takashi Nomura, Minoru Kubo, Jiwang Yan

  Japan Society of Mechanical Engineers, 2018, Journal of Advanced Mechanical Design, Systems and Manufacturing, 12(1) (1), JAMDSM0021, English

  [Refereed]

  Scientific journal


• Capturing an initial intermediate during the P450nor enzymatic reaction using time-resolved XFEL crystallography and caged-substrate

  Takehiko Tosha, Takashi Nomura, Takuma Nishida, Naoya Saeki, Kouta Okubayashi, Raika Yamagiwa, Michihiro Sugahara, Takanori Nakane, Keitaro Yamashita, Kunio Hirata, Go Ueno, Tetsunari Kimura, Tamao Hisano, Kazumasa Muramoto, Hitomi Sawai, Hanae Takeda, Eiichi Mizohata, Ayumi Yamashita, Yusuke Kanematsu, Yu Takano, Eriko Nango, Rie Tanaka, Osamu Nureki, Osami Shoji, Yuka Ikemoto, Hironori Murakami, Shigeki Owada, Kensuke Tono, Makina Yabashi, Masaki Yamamoto, Hideo Ago, So Iwata, Hiroshi Sugimoto, Yoshitsugu Shiro, Minoru Kubo

  Springer Science and Business Media LLC, Dec. 2017, Nature Communications, 8(1) (1), 1585

  [Refereed]

  Scientific journal

  Link to Kobe Univ. Repository (Kernel)


• Nanosecond pump-probe device for time-resolved serial femtosecond crystallography developed at SACLA

  Minoru Kubo, Eriko Nango, Kensuke Tono, Tetsunari Kimura, Shigeki Owada, Changyong Song, Fumitaka Mafuné, Ken Miyajima, Yoshihiro Takeda, Jun-Ya Kohno, Naoya Miyauchi, Takanori Nakane, Tomoyuki Tanaka, Takashi Nomura, Jan Davidsson, Rie Tanaka, Michio Murata, Takashi Kameshima, Takaki Hatsui, Yasumasa Joti, Richard Neutze, Makina Yabashi, So Iwata

  International Union of Crystallography, Sep. 2017, Journal of Synchrotron Radiation, 24(5) (5), 1086 - 1091, English

  Scientific journal

  Link to Kobe Univ. Repository (Kernel)


• Nanosecond pump-probe device for time-resolved serial femtosecond crystallography developed at SACLA

  Minoru Kubo, Eriko Nango, Kensuke Tono, Tetsunari Kimura, Shigeki Owada, Changyong Song, Fumitaka Mafune, Ken Miyajima, Yoshihiro Takeda, Jun-ya Kohno, Naoya Miyauchi, Takanori Nakane, Tomoyuki Tanaka, Takashi Nomura, Jan Davidsson, Rie Tanaka, Michio Murata, Takashi Kameshima, Takaki Hatsui, Yasumasa Joti, Richard Neutze, Makina Yabashi, So Iwata

  Sep. 2017, JOURNAL OF SYNCHROTRON RADIATION, 24, 1086 - 1091, English

  [Refereed]

  Scientific journal

  Link to Kobe Univ. Repository (Kernel)


• The Impact of the Polymer Chain Length on the Catalytic Activity of Poly(N-vinyl-2-pyrrolidone)-supported Gold Nanoclusters

  Setsiri Haesuwannakij, Tetsunari Kimura, Yuji Furutani, Kazu Okumura, Ken Kokubo, Takao Sakata, Hidehiro Yasuda, Yumi Yakiyama, Hidehiro Sakurai

  Aug. 2017, SCIENTIFIC REPORTS, 7(1) (1), 9579, English

  [Refereed]

  Scientific journal

  Link to Kobe Univ. Repository (Kernel)


• A nanosecond time-resolved XFEL analysis of structural changes associated with CO release from cytochrome c oxidase

  Atsuhiro Shimada, Minoru Kubo, Seiki Baba, Keitaro Yamashita, Kunio Hirata, Go Ueno, Takashi Nomura, Tetsunari Kimura, Kyoko Shinzawa-Itoh, Junpei Baba, Keita Hatano, Yuki Eto, Akari Miyamoto, Hironori Murakami, Takashi Kumasaka, Shigeki Owada, Kensuke Tono, Makina Yabashi, Yoshihiro Yamaguchi, Sachiko Yanagisawa, Miyuki Sakaguchi, Takashi Ogura, Ryo Komiya, Jiwang Yan, Eiki Yamashita, Masaki Yamamoto, Hideo Ago, Shinya Yoshikawa, Tomitake Tsukihara

  Jul. 2017, SCIENCE ADVANCES, 3(7) (7), e1603042, English

  [Refereed]

  Scientific journal

  Link to Kobe Univ. Repository (Kernel)


• Light-induced structural changes and the site of O=O bond formation in PSII caught by XFEL

  Michihiro Suga, Fusamichi Akita, Michihiro Sugahara, Minoru Kubo, Yoshiki Nakajima, Takanori Nakane, Keitaro Yamashita, Yasufumi Umena, Makoto Nakabayashi, Takahiro Yamane, Takamitsu Nakano, Mamoru Suzuki, Tetsuya Masuda, Shigeyuki Inoue, Tetsunari Kimura, Takashi Nomura, Shinichiro Yonekura, Long-Jiang Yu, Tomohiro Sakamoto, Taiki Motomura, Jing-Hua Chen, Yuki Kato, Takumi Noguchi, Kensuke Tono, Yasumasa Joti, Takashi Kameshima, Takaki Hatsui, Eriko Nango, Rie Tanaka, Hisashi Naitow, Yoshinori Matsuura, Ayumi Yamashita, Masaki Yamamoto, Osamu Nureki, Makina Yabashi, Tetsuya Ishikawa, So Iwata, Jian-Ren Shen

  Mar. 2017, NATURE, 543(7643) (7643), 131 - +, English

  [Refereed]

  Scientific journal


• A three-dimensional movie of structural changes in bacteriorhodopsin

  Eriko Nango, Antoine Royant, Minoru Kubo, Takanori Nakane, Cecilia Wickstrand, Tetsunari Kimura, Tomoyuki Tanaka, Kensuke Tono, Changyong Song, Rie Tanaka, Toshi Arima, Ayumi Yamashita, Jun Kobayashi, Toshiaki Hosaka, Eiichi Mizohata, Przemyslaw Nogly, Michihiro Sugahara, Daewoong Nam, Takashi Nomura, Tatsuro Shimamura, Dohyun Im, Takaaki Fujiwara, Yasuaki Yamanaka, Byeonghyun Jeon, Tomohiro Nishizawa, Kazumasa Oda, Masahiro Fukuda, Rebecka Andersson, Petra Bath, Robert Dods, Jan Davidsson, Shigeru Matsuoka, Satoshi Kawatake, Michio Murata, Osamu Nureki, Shigeki Owada, Takashi Kameshima, Takaki Hatsui, Yasumasa Joti, Gebhard Schertler, Makina Yabashi, Ana-Nicoleta Bondar, Jorg Standfuss, Richard Neutze, So Iwata

  Dec. 2016, SCIENCE, 354(6319) (6319), 1552 - 1557, English

  [Refereed]

  Scientific journal


• Lipidic cubic phase injector is a viable crystal delivery system for time-resolved serial crystallography

  Przemyslaw Nogly, Valerie Panneels, Garrett Nelson, Cornelius Gati, Tetsunari Kimura, Christopher Milne, Despina Milathianaki, Minoru Kubo, Wenting Wu, Chelsie Conrad, Jesse Coe, Richard Bean, Yun Zhao, Petra Bath, Robert Dods, Rajiv Harimoorthy, Kenneth R. Beyerlein, Jan Rheinberger, Daniel James, Daniel DePonte, Chufeng Li, Leonardo Sala, Garth J. Williams, Mark S. Hunter, Jason E. Koglin, Peter Berntsen, Eriko Nango, So Iwata, Henry N. Chapman, Petra Fromme, Matthias Frank, Rafael Abela, Sebastien Boutet, Anton Barty, Thomas A. White, Uwe Weierstall, John Spence, Richard Neutze, Gebhard Schertler, Jorg Standfuss

  Aug. 2016, NATURE COMMUNICATIONS, 7, 12314, English

  [Refereed]

  Scientific journal


• A Study of the Dynamics of the Heme Pocket and C-helix in CooA upon CO Dissociation Using Time-Resolved Visible and UV Resonance Raman Spectroscopy

  Akihiro Otomo, Haruto Ishikawa, Misao Mizuno, Tetsunari Kimura, Minoru Kubo, Yoshitsugu Shiro, Shigetoshi Aono, Yasuhisa Mizutani

  Aug. 2016, JOURNAL OF PHYSICAL CHEMISTRY B, 120(32) (32), 7836 - 7843, English

  [Refereed]

  Scientific journal


• A nearly on-axis spectroscopic system for simultaneously measuring UV-visible absorption and X-ray diffraction in the SPring-8 structural genomics beamline

  Miyuki Sakaguchi, Tetsunari Kimura, Takuma Nishida, Takehiko Tosha, Hiroshi Sugimoto, Yoshihiro Yamaguchi, Sachiko Yanagisawa, Go Ueno, Hironori Murakami, Hideo Ago, Masaki Yamamoto, Takashi Ogura, Yoshitsugu Shiro, Minoru Kubo

  Jan. 2016, JOURNAL OF SYNCHROTRON RADIATION, 23, 334 - 338, English

  [Refereed]

  Scientific journal


• Ultraprecision cutting of single-crystal calcium fluoride for fabricating micro flow cell for protein structure analysis

  Ryo Komiya, Tetsunari Kimura, Minoru Kubo, Jiwang Yan

  Japan Society of Mechanical Engineers, Oct. 2015, Proceedings of the 8th International Conference on Leading Edge Manufacturing in 21st Century, LEM 2015, English

  International conference proceedings


• Chimeras of Channelrhodopsin-1 and-2 from Chlamydomonas reinhardtii Exhibit Distinctive Light-induced Structural Changes from Channelrhodopsin-2

  Asumi Inaguma, Hisao Tsukamoto, Hideaki E. Kato, Tetsunari Kimura, Toru Ishizuka, Satomi Oishi, Hiromu Yawo, Osamu Nureki, Yuji Furutani

  May 2015, JOURNAL OF BIOLOGICAL CHEMISTRY, 290(18) (18), 11623 - 11634, English

  [Refereed]

  Scientific journal


• New insights into metal ion-crown ether complexes revealed by SEIRA spectroscopy

  Yoshiya Inokuchi, Takayuki Ebata, Toshiaki Ikeda, Takeharu Haino, Tetsunari Kimura, Hao Guo, Yuji Furutani

  2015, NEW JOURNAL OF CHEMISTRY, 39(11) (11), 8673 - 8680, English

  [Refereed]

  Scientific journal


• Formation of host-guest complexes on gold surface investigated by surface-enhanced IR absorption spectroscopy

  Yoshiya Inokuchi, Takahiro Mizuuchi, Takayuki Ebata, Toshiaki Ikeda, Takeharu Haino, Tetsunari Kimura, Hao Guo, Yuji Furutani

  Jan. 2014, Chemical Physics Letters, 592, 90 - 95, English

  [Refereed]

  Scientific journal


• 3P092 Molecular mechanism for substrate NO binding to bacterial Nitric Oxide Reductase(02. Heme proteins,Poster,The 52nd Annual Meeting of the Biophysical Society of Japan(BSJ2014))

  Ishii Shoko, Kimura Tetsunari, Tosha Takehiko, Shiro Yoshitsugu, Kubo Minoru

  The Biophysical Society of Japan General Incorporated Association, 2014, Seibutsu Butsuri, 54(1) (1), S264, English


• 2P092 Characterization of transient NO-bound form of Nitric Oxide Reductase by Transient Absorption Spectroscopy(02. Heme proteins,Poster,The 52nd Annual Meeting of the Biophysical Society of Japan(BSJ2014))

  Kimura Tetsunari, Ishii Shoko, Tosha Takehiko, Shiro Yoshitsugu, Kubo Minoru

  The Biophysical Society of Japan General Incorporated Association, 2014, Seibutsu Butsuri, 54(1) (1), S210, English


• Distortion of the amide-I and -II bands of an alpha-helical membrane protein, pharaonis halorhodopsin, depends on thickness of gold films utilized for surface-enhanced infrared absorption spectroscopy

  Hao Guo, Tetsunari Kimura, Yuji Furutani

  Jun. 2013, CHEMICAL PHYSICS, 419, 8 - 16, English

  [Refereed]

  Scientific journal


• 1P241 Comparative analysis of Channelrhodopsin and its chimeras based on FTIR spectroscopy(18A. Photobiology: Vision & Photoreception,Poster)

  Inaguma Asumi, Tsukamoto Hisao, Kimura Tetsunari, Ishizuka Toru, Yawo Hiromu, Furutani Yuji

  The Biophysical Society of Japan General Incorporated Association, 2013, Seibutsu Butsuri, 53(1) (1), S145, English


• 1P097 Development of micro-channel flow-flash method for time-resolved spectroscopic study of enzymatic reactions(02.Heme proteins,Poster,The 51st Annual Meeting of the Biophysical Society of Japan)

  Kimura Tetsunari, Tosha Takehiko, Shiro Yoshitsugu, Kubo Minoru

  The Biophysical Society of Japan General Incorporated Association, 2013, Seibutsu Butsuri, 53(1) (1), S122, English


• 1P221 Development of a Rapid Buffer-Exchange System for Time-Resolved ATR-FTIR Spectroscopy with the Step-Scan Mode(13C. Biological & Artifical membrane: Excitation & Channels,Poster)

  Furutani Yuji, Kimura Tetsunari, Okamoto Kido

  The Biophysical Society of Japan General Incorporated Association, 2013, Seibutsu Butsuri, 53(1) (1), S142, English


• Development of a rapid Buffer-exchange system for time-resolved ATR-FTIR spectroscopy with the step-scan mode

  Yuji Furutani, Tetsunari Kimura, Kido Okamoto

  2013, Biophysics (Japan), 9, 123 - 129, English

  [Refereed]

  Scientific journal


• Dynamics of Dangling Bonds of Water Molecules in pharaonis Halorhodopsin during Chloride Ion Transportation

  Yuji Furutani, Kuniyo Fujiwara, Tetsunari Kimura, Takashi Kikukawa, Makoto Demura, Hideki Kandori

  Oct. 2012, JOURNAL OF PHYSICAL CHEMISTRY LETTERS, 3(20) (20), 2964 - 2969, English

  [Refereed]

  Scientific journal


• 1C1558 Development of novel rapid buffer-exchange system and microfluidic mixer for time-resolved spectroscopic study of low-yield biomolecules(Proteins: Measurement, Analysis, Engineering,Oral Presentation,The 50th Annual Meeting of the Biophysical Society of Japan)

  Kimura Tetsunari, Furutani Yuji

  The Biophysical Society of Japan General Incorporated Association, 2012, Seibutsu Butsuri, 52, S24, English


• 2PT182 Water hydrogen-bonding dynamics in the function of pharaonis halorhodopsin studied by time-resolved FTIR spectroscopy(The 50th Annual Meeting of the Biophysical Society of Japan)

  Fujiwara Kuniyo, Kimura Tetsunari, Kikukawa Takashi, Demura Makoto, Kandori Hideki, Furutani Yuji

  The Biophysical Society of Japan General Incorporated Association, 2012, Seibutsu Butsuri, 52, S136 - S137, English


• 2PT183 Structural changes in the chimeric Channelrhodopsins revealed by ATR-FTIR spectroscopy(The 50th Annual Meeting of the Biophysical Society of Japan)

  Inaguma Asumi, Tsukamoto Hisao, Kimura Tetsunari, Ishizuka Toru, Yawo Hiromu, Furutani Yuji

  The Biophysical Society of Japan General Incorporated Association, 2012, Seibutsu Butsuri, 52, S137, English


• Direct visualization reveals dynamics of a transient intermediate during protein assembly

  Xin Zhang, Vinh Q. Lam, Yun Mou, Tetsunari Kimura, Jaeyoon Chung, Sowmya Chandrasekar, Jay R. Winkler, Stephen L. Mayo, Shu-ou Shan

  Apr. 2011, PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 108(16) (16), 6450 - 6455, English

  [Refereed]

  Scientific journal


• Time-resolved small-angle X-ray scattering study of the folding dynamics of barnase

  Tsuyoshi Konuma, Tetsunari Kimura, Shuzo Matsumoto, Yuji Goto, Tetsuro Fujisawa, Alan R. Fersht, Satoshi Takahashi

  Feb. 2011, Journal of Molecular Biology, 405(5) (5), 1284 - 1294, English

  [Refereed]

  Scientific journal


• 2SH-02 Development of Stopped-flow Attenuated Total Reflection FTIR Spectroscopy for Detecting Dynamic Strucutral Changes in Ion Transporters(2SH New Experimental Tools for Structural Changes of Membrane Proteins: Beyond X-ray Structures,The 49th Annual Meeting of the Biophysical Society of Japan)

  Furutani Yuji, Kimura Tetsunari

  The Biophysical Society of Japan General Incorporated Association, 2011, Seibutsu Butsuri, 51, S20, English


• 2Q1624 Preparation and Characterization of the Fusion Protein : KcsA and Microbial Rhodopsins(Photobiology: Vision & Photoreception 2,The 48th Annual Meeting of the Biophysical Society of Japan)

  Muramatsu Chikako, Kawanabe Akira, Kimura Tetsunari, Kandori Hideki, Furutani Yuji

  The Biophysical Society of Japan General Incorporated Association, 2011, Seibutsu Butsuri, 51, S103, English


• 3B1024 Ion-selective mechanism of Mg^<2+> transporter MgtE studied by attenuated total refrection FTIR spectroscopy(3B Membrane proteins 2,The 49th Annual Meeting of the Biophysical Society of Japan)

  Kimura Tetsunari, Douki Shintaro, Ishitani Ryuichiro, Nureki Osamu, Furutani Yuji

  The Biophysical Society of Japan General Incorporated Association, 2011, Seibutsu Butsuri, 51, S109, English


• 3A1546 SEIRA spectroscopy of a halorhodopsin monolayer on gold island film(3A Biol & Artifi memb 4: Transport, Signal transduction,The 49th Annual Meeting of the Biophysical Society of Japan)

  Guo Hao, Kimura Tetsunari, Furutani Yuji

  The Biophysical Society of Japan General Incorporated Association, 2011, Seibutsu Butsuri, 51, S108, English


• 2P031 1G1505 Conformational changes upon chloride-ion pumping during the pharaonis halorhodopsin photocycle studied by time-resolved FTIR spectroscopy(The 48th Annual Meeting of the Biophysical Society of Japan)

  Kimura Tetsunari, Kandori Hideki, Furutani Yuji

  The Biophysical Society of Japan General Incorporated Association, 2010, Seibutsu Butsuri, 50(2) (2), S87, English


• 2SC1350 In vitro protein folding dynamics in the microsecond to millisecond timescale(2SC Advanced Insight into Protein Motions-in silico, in vitro and in vivo Studies,The 48th Annual Meeting of the Biophysical Society of Japan)

  Kimura Tetsunari

  The Biophysical Society of Japan General Incorporated Association, 2010, Seibutsu Butsuri, 50(2) (2), S11, English


• Dehydration of main-chain amides in the final folding step of single-chain monellin revealed by time-resolved infrared spectroscopy

  Tetsunari Kimura, Akio Maeda, Shingo Nishiguchi, Koichiro Ishimori, Isao Morishima, Takashi Konno, Yuji Goto, Satoshi Takahashi

  Sep. 2008, PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 105(36) (36), 13391 - 13396, English

  [Refereed]

  Scientific journal


• 2P078 Time-resolved small angle x-ray scattering study on the burst phase intermediate in the folding of bamase(Proteins-stability, folding, and other physicochemical properties,Poster Presentations)

  Konuma Tsuyoshi, Kimura Tetsunari, Matsumoto Shuzo, Goto Yuji, Fujisawa Tetsuro, Takahashi Satoshi

  The Biophysical Society of Japan General Incorporated Association, 2007, Seibutsu Butsuri, 47, S132, English


• Site-specific collapse dynamics guide the formation of the cytochrome c ' four-helix bundle

  Tetsunari Kimura, Jennifer C. Lee, Harry B. Gray, Jay R. Winkler

  Jan. 2007, PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 104(1) (1), 117 - 122, English

  [Refereed]

  Scientific journal


• Time-resolved small-angle X-ray scattering investigation of the folding dynamics of heme oxygenase: Implication of the scaling relationship for the submillisecond intermediates of protein folding

  T Uzawa, T Kimura, K Ishimori, Morishima, I, T Matsui, M Ikeda-Saito, S Takahashi, S Akiyama, T Fujisawa

  Mar. 2006, JOURNAL OF MOLECULAR BIOLOGY, 357(3) (3), 997 - 1008, English

  [Refereed]

  Scientific journal


• S3f1-4 Generality of Initial Collapse Demonstrated by Scaling Relationship for Submillisecond Intermediates of Protein Folding(S3-f1: "Hydration Effects on Structure and Thermodynamics of Proteins,Symposia,Abstract,Meeting Program of EABS & BSJ 2006)

  Uzawa Takanori, Kimura Tetsunari, Ishimori Koichiro, Morishima Isao, Matsui Toshitaka, Ikeda-Saito Masao, Takahashi Satoshi, Akiyama Shuji, Fujisawa Tetsuro

  The Biophysical Society of Japan General Incorporated Association, 2006, Seibutsu Butsuri, 46(2) (2), S139, English


• Dehydration in the folding of reduced cytochrome c revealed by the electron-transfer-triggered folding under high pressure

  T Kimura, K Sakamoto, Morishima, I, K Ishimori

  Jan. 2006, JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 128(3) (3), 670 - 671, English

  [Refereed]

  Scientific journal


• Specifically collapsed intermediate in the early stage of the folding of ribonuclease A

  Tetsunari Kimura, Shuji Akiyama, Takanori Uzawa, Koichiro Ishimori, Isao Morishima, Tetsuro Fujisawa, Satoshi Takahashi

  Jul. 2005, Journal of Molecular Biology, 350(2) (2), 349 - 362, English

  [Refereed]

  Scientific journal


• Specific collapsed followed by slow hydrogen-bond formation of β-sheet in the folding of single-chain monellin

  Tetsunari Kimura, Takanori Uzawa, Koichiro Ishimori, Isao Morishima, Satoshi Takahashi, Takashi Konno, Shuji Akiyama, Tetsuro Fujisawa

  Feb. 2005, Proceedings of the National Academy of Sciences of the United States of America, 102(8) (8), 2748 - 2753, English

  [Refereed]

  Scientific journal


• Collapse and search dynamics of apomyoglobin folding revealed by submillisecond observations of α-helical content and compactness

  Takanori Uzawa, Shuji Akiyama, Tetsunari Kimura, Satoshi Takahashi, Koichiro Ishimori, Isao Morishima, Tetsuro Fujisawa

  Feb. 2004, Proceedings of the National Academy of Sciences of the United States of America, 101(5) (5), 1171 - 1176, English

  [Refereed]

  Scientific journal


• 3P023 Significance of oligomerization and rapid collapse in hemeoxygenase folding

  Uzawa T., Kimura T., Takahashi S., Ishimori K., Akiyama S., Fujisawa T., Matsui T., Saito M.

  The Biophysical Society of Japan General Incorporated Association, 2004, Seibutsu Butsuri, 44, S195, Japanese


• 3P017 Contribution of dehydration to reduced cytochrome c folding revealed by high pressure measurements

  Kimura T., Sekiyama N., Sakamoto K., Morishima I., Ishimori K.

  The Biophysical Society of Japan General Incorporated Association, 2004, Seibutsu Butsuri, 44, S194, Japanese


• Main-chain collapse and subsequent tertiary structure formation during apomyoglobin folding

  Uzawa T., Kimura T., Takahashi S., Ishimori K., Morishima I., Akiyama S., Fujisawa T.

  The Biophysical Society of Japan General Incorporated Association, 2003, Seibutsu Butsuri, 43, S60, Japanese


• Roles of the N-terminal fluctuating domain in folding of coiled coil GCN4-pl'

  Kinoshita M., Kimura T., Takahashi S., Ishimori K., Morishima I.

  The Biophysical Society of Japan General Incorporated Association, 2003, Seibutsu Butsuri, 43, S61, Japanese


• Folding Mechanism of Single-chain Monellin Based on the Secondary Structure Formation and the Packing of the Hydrophobic Residues

  Kimura T., Takahashi S., Konno T., Ishimori K., Morishima I.

  The Biophysical Society of Japan General Incorporated Association, 2003, Seibutsu Butsuri, 43, S59, Japanese


• Conformational landscape of cytochrome c folding studied by microsecond-resolved small-angle x-ray scattering

  Shuji Akiyama, Satoshi Takahashi, Tetsunari Kimura, Koichiro Ishimori, Isao Morishima, Yukihiro Nishikawa, Tetsuro Fujisawa

  Feb. 2002, Proceedings of the National Academy of Sciences of the United States of America, 99(3) (3), 1329 - 1334, English

  [Refereed]

  Scientific journal


• 3M1030 Observation on the folding process of single-chain monellin studied by microsecond-resolved FTIR spectroscopy

  Kimura T., Takahashi S., Konno T., Ishimori K., Morishima I.

  The Biophysical Society of Japan General Incorporated Association, 2002, Seibutsu Butsuri, 42(2) (2), S190, Japanese


• Apomyoglobin folding studied by submillisecond small-angle X-ray scattering

  Uzawa T, Akiyama S, Kimura T, Takahashi S, Ishimori K, Morishima I, Nishikawa Y, Fujisawa T

  The Biophysical Society of Japan General Incorporated Association, 2001, Seibutsu Butsuri, 41, S166, Japanese


• The study on the burst phase of ribonuclease A folding using a rapid mixing device

  Kimura T., Akiyama S., Takahashi S., Harada Y., Ishimori K., Morishima I.

  The Biophysical Society of Japan General Incorporated Association, 2000, Seibutsu Butsuri, 40, S161, Japanese

• ドデシル硫酸ナトリウム共存下でのαシヌクレイン凝集反応の多様性

  北野さくら, 柚佳祐, 笹田航, 木村哲就, 茶谷絵理

  2023, 日本蛋白質科学会年会プログラム・要旨集, 23rd (CD-ROM)


• 鉄還元機能を持つ6回膜貫通型タンパク質101F6の大腸菌での発現と機能解析

  阪口智哉, 澤井仁美, 城宜嗣, 鍔木基成, 當舎武彦, 木村哲就, 杉本宏

  2023, 日本生化学会大会(Web), 96th


• 二液混合シリアルフェムト秒結晶構造解析による酵素反応追跡

  南後恵理子, 南後恵理子, LUO Fangjia, LUO Fangjia, 木村哲就, 菅原道泰, 中根崇智, 鈴木守, 桝田哲哉, 溝端栄一, 登野健介, 登野健介, 岩田想, 岩田想

  2021, 日本結晶学会年会講演要旨集, 2021


• Molecular Mechanism of the Catalytic Reaction of no Reductase Revealed by Novel Time-Resolved Visible/IR Absorption Spectrometers with Microfluidic Device

  Tetsunari Kimura, Hanae Takeda, Shoko Ishii, Takehiko Tosha, Yoshitsugu Shiro, Minoru Kubo

  Feb. 2016, BIOPHYSICAL JOURNAL, 110(3) (3), 548A - 548A, English

  Summary international conference


• SACLAにおけるシリアルフェムト秒X線結晶構造解析の技術開発

  田中智之, 菅原道泰, 中根崇智, 木村哲就, 木村哲就, 久保稔, 久保稔, 宮島謙, 宮島謙, 登野健介, 真船文隆, 真船文隆, 南後恵理子, 南後恵理子, 岩田想, 岩田想

  2016, 日本結晶学会年会講演要旨集, 2016


• 時分割シリアルフェムト秒X線結晶解析法によるバクテリオロドプシン光反応中間体の構造解析

  南後恵理子, NEUTZE Richard, 久保稔, 久保稔, 木村哲就, 中根崇智, ROYANT Antoine, 登野健介, 岩田想

  2015, 日本結晶学会年会講演要旨集, 2015


• 急速緩衝液交換法による時間分解全反射赤外分光法の開発

  古谷 祐詞, 木村 哲就, 岡本 基土

  日本生物物理学会, 30 Sep. 2014, 生物物理, 54(5) (5), 272 - 275, Japanese

  [Refereed][Invited]

  Introduction scientific journal


• C-2 霊長類、視覚・味覚のGPCR型受容体に対する赤外分光研究

  神取 秀樹, 片山 耕大, 古谷 祐詞, 木村 哲就

  京都大学霊長類研究所, 04 Oct. 2012, 霊長類研究所年報, 42, 118 - 118, Japanese


• Conformational changes upon chloride-ion pumping during the pharaonis halorhodopsin photocycle studied by time-resolved FTIR difference spectroscopy

  Tetsunari Kimura, Hideki Kandori, Yuji Furutani

  Mar. 2011, ABSTRACTS OF PAPERS OF THE AMERICAN CHEMICAL SOCIETY, 241, English

  Summary international conference


• PHYS 341-Folding dynamics of acyl-CoA binding protein probed by energy-transfer kinetics measurements

  Tetsunari Kimura, Harry B. Gray, Jay R. Winkler

  Aug. 2008, ABSTRACTS OF PAPERS OF THE AMERICAN CHEMICAL SOCIETY, 236, English

  Summary international conference


• Direct observation of the multistep helix formation of poly-L-glutamic acids using microsecond-resolved FTIR and CD spectroscopies.

  T Kimura, S Takahashi, S Akiyama, T Uzawa, K Ishimori, Morishima, I

  Feb. 2003, BIOPHYSICAL JOURNAL, 84(2) (2), 162A - 162A, English

  Summary international conference


• Direct observation of the multistep helix formation of poly-L-glutamic acids

  T Kimura, S Takahashi, S Akiyama, T Uzawa, K Ishimori, Morishima, I

  Oct. 2002, JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 124(39) (39), 11596 - 11597, English

• ヘムタンパク質の科学 : 生理機能の理解とその展開に向けて

  城, 宜嗣, 青野, 重利, 齋藤, 正男

  Contributor, 第１編 第５章 第５節 反応速度論, エヌ・ティー・エス, May 2022, Japanese, ISBN: 9784860437787

• ABCトランスポーターにおける基質輸送の分光学的解析

  Tetsunari Kimura

  第7回広帯域極限電磁波生命理工連携研究会 「生命現象に関わる分子構造ダイナミクス解析の最新」, Jan. 2025, Japanese, Domestic conference

  [Invited]

  Oral presentation


• Spectroscopy in heme transport of the ABC transporter; BhuUV-T

  中川開斗, 城宜嗣, 杉本宏, 木村哲就

  第36回生物無機化学夏季セミナー第4回生命金属科学夏合宿合同大会, Sep. 2024, Japanese, Domestic conference

  Poster presentation


• Coupling of ATP reactions with allocrite transport in heme ABC transporter; BhuUV-T, revealed by time-resolved spectroscopy.

  Tetsunari Kimura, Ayaka Naka, Akiho Hara, Yasuhiro Kobori, Yoshitsugu Shiro, Hiroshi Sugimoto

  IUPAB2024, Jun. 2024, English, International conference

  Poster presentation


• 部位特異的スピンラベルによる ABC トランスポーター BhuUV-T の輸送機構の解明

  炭田悠貴, 仲絢香, 原明穂, 小堀康博, 城宜嗣, 杉本宏, 木村 哲就

  第50回生体分子科学討論会, Jun. 2024, Japanese, 第50回生体分子科学討論会・第3回生命金属科学シンポジウム実行委員会, 慶應義塾大学日吉キャンパス, Domestic conference

  Poster presentation


• シトクロムb561D2の機能性評価に資する発現・精製系の構築

  吉村悠, 鍔木基成, 杉本宏, 木村哲就

  第50回生体分子科学討論会, Jun. 2024, Japanese, 第50回生体分子科学討論会・第3回生命金属科学シンポジウム実行委員会, 慶應義塾大学日吉キャンパス, Domestic conference

  Poster presentation


• ABCトランスポーターBhuUV-Tのヘム輸送における初期段階の紫外・可視吸収分光解析

  中川開斗, 城宜嗣, 杉本宏, 木村哲就

  第50回生体分子科学討論会, Jun. 2024, Japanese, Domestic conference

  Poster presentation


• UV/visible absorption spectroscopy of the early steps in heme transport of the ABC transporter; BhuUV-T

  Kaito Nakagawa, Yoshitsugu, Hiroshi Sugimoto, Tetsunari Kimura

  The 24th Annual Meeting of the Protein Science Society of Japan, Jun. 2024, English, Domestic conference

  Poster presentation


• Spectroscopic analysis of BhuUV-T reconstituted with amphiphilic polymers

  Yuki Sumida, Ayaka Naka, Yasuhiro Kobori, Yoshitsugu Shiro, Hiroshi Sugimoto, Tetsunari Kimura

  The 61st Annual Meeting of the Biophysical Society of Japa, Nov. 2023, English, 名古屋国際会議場, Domestic conference

  Poster presentation


• Thermodynamic and kinetic analyses of interaction between aequorin and Ca2+

  Urara Kuroki, Yuka Osaki, Toshiya Funahashi, Toru Nakatsu, Tetsunari Kimura

  第96回日本生化学会大会, Nov. 2023, Japanese, 福岡国際会議場, Domestic conference

  Poster presentation


• Investigation of the mechanism of heme transport in the ABC transporter BhuUV-T by spectroscopic methods

  Akiho Hara, Yoshitsugu Shiro, Hiroshi Sugimoto, Tetsunari Kimura

  第96回日本生化学会大会, Nov. 2023, Japanese, 福岡国際会議場, Domestic conference

  Poster presentation


• 鉄還元機能を持つ6回膜貫通型タンパク質101F6の大腸菌での発現と機能解析

  阪口 智哉, 澤井 仁美, 城 宜嗣, 鍔木 基成, 當舎 武彦, 木村 哲就, 杉本 宏

  第96回日本生化学会大会, Nov. 2023, Japanese, 福岡国際会議場, Domestic conference

  Poster presentation


• Development of microfluidic mixers for time-resolved SFX and spectroscopy and their applications

  Tetsunari KIMURA

  UK/Japan meeting and workshop “Dynamic Crystallography-XFELs and Synchrotrons to study enzyme reactions”, Sep. 2023, English, University of Lancaster, International conference

  [Invited]

  Oral presentation


• Time-resolved Measurements for Structural and Functional Dynamics of Membrane Protein Complexes

  Tetsunari Kimura, Ayaka Naka, Akiho Hara, Yoshitsugu Shiro, Hiroshi Sugimoto

  The 9th International Discussion Meeting on Relaxations in Complex Systems (9 IDMRCS), Aug. 2023, English, Makuhari Messe, International conference

  [Invited]

  Oral presentation


• Thermodynamic and kinetic analyses of chemical luminescence induced by the Ca2+ binding to Aequorin

  Urara Kuroki, Yuka Osaki, Toshiya Funahashi, Toru Nakatsu, Tetsunari Kimura

  The 23nd Annual Meeting of the Protein Science Society of Japan, Jul. 2023, English, 名古屋国際会議場, Domestic conference

  Poster presentation


• Time-resolved spectroscopy of heme transport in ABC transporter; BhuUV-T

  Akiho Hara, Yoshitsugu Shiro, Hiroshi Sugimoto, Tetsunari Kimura

  The 23nd Annual Meeting of the Protein Science Society of Japan, Jul. 2023, English, 名古屋国際会議場, Domestic conference

  Poster presentation


• Thermodynamic and kinetic analyses of interaction between aequorin and Ca2+

  木村哲就, 大崎優香, 黒木麗, 船橋俊也, 中津亨

  新学術領域研究「高速分子動画」領域セミナー, May 2023, English, RIKEN, Domestic conference

  Poster presentation


• 時間分解分光測定によるタンパク質の機能・構造解析

  Tetsunari KIMURA

  東北大学多元研セミナー, Feb. 2023, Japanese, 東北大多元研, 東北大多元研, Domestic conference

  [Invited]

  Oral presentation


• 物理化学的解析による発光タンパク質イクオリンの反応機構

  木村哲就, 大崎優香, 黒木麗, 船橋俊也, 中津亨

  新学術領域研究「高速分子動画」領域会議, Nov. 2022, Japanese, MEXT Grant-in-Aid for Scientific Research on Innovative Area ”Non-equilibrium-state molecular movies and their applications (Molecular Movies)", 淡路夢舞台, Domestic conference

  Poster presentation


• Development and application of complementary spectroscopic methods for dynamic structural biology

  Tetsunari KIMURA

  IPR seminar, Oct. 2022, English, "Cooperation: RIKEN Pioneering Project ""Biology of Intracellular Environments"" MEXT Program for Promoting Researches on the Supercomputer Fugaku ""Biomolecular dynamics in a living cell"" MEXT Grant-in-Aid for Transformative Research Areas (A) ""New Cross Scale Biology"" MEXT Grant-in-Aid for Scientific Research on Innovative Area ”Non-equilibrium-state molecular movies and their applications (Molecular Movies)", Institute for Protein Science, Osaka Univ., International conference

  [Invited]

  Oral presentation


• Thermodynamic analysis of the calcium binding with photoluminescence protein; aequorin

  Kuroki Urara, Funahashi Toshiya, Onishi Yusuke, Nakatsu Toru, Kimura Tetsunari

  第60回日本生物物理学会年会, Sep. 2022, English, 函館アリーナ・函館市民会館, Domestic conference

  Poster presentation


• Conformational changes in transmembrane domain of ABC transporter revealed by double spin label-ESR spectroscopy.

  Naka Ayaka、Kobori Yasuhiro、Tsubaki Motonari、Shiro Yoshitsugu、Sugimoto Hiroshi、Kimura Tetsunari

  第60回日本生物物理学会年会, Sep. 2022, English, 函館アリーナ・函館市民会館, Domestic conference

  Poster presentation


• Tire-resolved spectroscopic analysis of allocrite transport mechanism in heme ABC transporter; BhuUV-T

  Akiho Hara, Yoshitsugu Shiro, Hiroshi Sugimoto, Tetsunari Kimura

  第60回日本生物物理学会年会, Sep. 2022, English, 函館アリーナ・函館市民会館, Domestic conference

  Poster presentation


• Analysis of lipid peroxidation induced by iron-reducing membrane heme protein; CYB561D2 in nanodiscs

  Aoi Yamaguchi, Motonari Tsubaki, Tetsunari Kimura

  第60回日本生物物理学会年会, Sep. 2022, English, 函館アリーナ・函館市民会館, Domestic conference

  Poster presentation


• ITCを用いたイクオリンCa2+間相互作用の熱力学的解析

  黒木麗, 船橋俊也, 大西裕介, 中津亨, 木村哲就

  第48回生体分子科学討論会, Jun. 2022, Japanese, とりぎん文化会館小ホール, Domestic conference

  Poster presentation


• 101F6ナノディスクによる電子移動反応の実験的解析

  Aoi Yamaguchi, Motonari Tsubaki, Tetsunari Kimura

  第48回生体分子科学討論会, Jun. 2022, Japanese, とりぎん文化会館小ホール, Domestic conference

  Poster presentation


• 二重スピンラベル-ESR分光法を用いた ABCトランスポーターBhuUV-Tの構造解析

  仲絢香, 小堀 康博, 鍔木 基成, 城 宜嗣, 杉本 宏, 木村哲就

  第48回生体分子討論会, Jun. 2022, Japanese, とりぎん文化会館小ホール, Domestic conference

  Poster presentation


• ABCトランスポーターBhuUV-Tにおけるヘム輸送機構の分光学的解析

  原明穂, 城宜嗣, 杉本宏, 木村哲就

  第48回生体分子科学討論会, Jun. 2022, Japanese, とりぎん文化会館小ホール, Domestic conference

  Poster presentation


• ヘムABCトランスポーター; BhuUV-T の膜輸送に関するキネティクス解析

  Tetsunari KIMURA

  第22回日本蛋白質科学会年会, Jun. 2022, English, つくば国際会議場, Domestic conference

  [Invited]

  Oral presentation


• Thermodynamic analysis of interaction between Aequorin and Ca2+ by ITC

  Urara Kuroki, Toshiya Funahashi, Yusuke Onishi, Toru Nakatsu, Tetsunari Kimura

  The 22nd Annual Meeting of the Protein Science Society of Japan, Jun. 2022, English, つくば国際会議場, Domestic conference

  Oral presentation


• Spectroscopic analysis of electron transfer reaction in iron-reducing membrane protein 101F6- nanodiscs

  Aoi Yamaguchi, Motonari Tsubaki, Tetsunari Kimura

  The 22nd annual Meeting of the Protein Sience Society of Japan, Jun. 2022, English, つくば国際会議場, Domestic conference

  Oral presentation


• ATP binding state of heme ABC transporter, BhuUV-T, investigated by double spin-labeling ESR spectroscopy

  Ayaka Naka, Yasuhiro Kobori, Motonari Tsubaki, Yositsugu Shiro, Hiroshi Sugimoto, Tetsunari Kimura

  the 22nd Annual Meeting og the Protein Science Society of Japan, Jun. 2022, English, つくば国際会議場, Domestic conference

  Oral presentation


• Time-resolved spectroscopic measurements on the mechanism of heme transport by the ABC transporter BhuUV-T

  Akiho Hara, Yoshitsugu Shiro, Hiroshi Sugimoto, Tetsunari Kimura

  第22回日本蛋白質科学会年会, Jun. 2022, English, つくば国際会議場, Domestic conference

  Oral presentation


• Microspectroscopic systemsfor time-resolved measurementsof protein microcrystals

  Tetsunari KIMURA

  新学術領域研究「高速分子動画」領域会議, May 2022, Japanese, MEXT Grant-in-Aid for Scientific Research on Innovative Area ”Non-equilibrium-state molecular movies and their applications (Molecular Movies)", RIKEN, Domestic conference

  [Invited]

  Oral presentation


• マイクロ流路デバイスを用いた時間分解顕微分光法による膜輸送の分子機構解析

  木村 哲就

  神戸大学先端融合研究環開拓プロジェクトシンポジウム, Mar. 2022, Japanese, Online, Domestic conference

  [Invited]

  Invited oral presentation


• 膜輸送タンパク質 ABC トランスポーターの機能・構造変化の分光学的解析

  木村 哲就

  日本分光学会 関西支部令和３年度講演会, Mar. 2022, Japanese, 日本分光学会 関西支部, Online, Domestic conference

  [Invited]

  Invited oral presentation


• Developments and applications of time-resolved microspectroscopy for protein micro-crystals

  Tetsunari Kimura

  PacifiChem2021, Dec. 2021, English, Pacific Basin Chemical Societies, Online, International conference

  Oral presentation


• Structural changes of a-synuclein, an intrinsically disordered protein, along the lipid-binding and oligomerization revealed by fluorescence lifetime measurements

  Ko Sasada, Tetsunari Kimura, Ryosuke Matsubara, Koichi Fujii

  PacifiChem2021, Dec. 2021, English, Pacific Basin Chemical Societies, Online, International conference

  Poster presentation


• Transmembrane electron-transfer reaction of 101F6 protein reconstituted into nanodisc

  Aoi Yamaguchi, Hamed A. Abosharaf, Motonari Tsubaki, Tetsunari Kimura

  PacifiChem2021, Dec. 2021, English, Pacific Basin Chemical Societies, Online, International conference, Co-authored internationally

  Poster presentation


• Spectroscopic analysis of electron transfer reaction in iron-reducing membrane protein 101F6 reconstituted into nanodiscs

  Aoi Yamaguchi, Hamed A. Abosharaf, Motonari Tsubaki, Tetsunari Kimura

  第59回日本生物物理学会年会, Nov. 2021, English, Online, Domestic conference, Co-authored internationally

  Oral presentation


• Structural analyses of ABC transporters in nucleotide bound states investigated by CW-ESR spectroscopy

  Ayaka Naka, Yasuhiro Kobori, Motonari Tsubaki, Yoshitsugu Shiro, Hiroshi Sugimoto, Tetsunari Kimura

  第59回日本生物物理学会年会, Nov. 2021, English, Online, Domestic conference

  Oral presentation


• Structural analysis of α-Synuclein in the lipid bilayer bound state by fluorescence lifetime measurements

  Ko Sasada, Ryosuke Matsubara, Koichi Fujii, Tetsunari Kimura

  第59回日本生物物理学会年会, Nov. 2021, English, Online, Domestic conference

  Oral presentation


• Kinetic analysis of the transport in heme ABC transporter; BhuUV-T, by time-resolved spectroscopy

  Tetsunari Kimura

  第59回日本生物物理学会年会, Nov. 2021, English, 日本生物物理学会, Online, Domestic conference

  [Invited]

  Invited oral presentation


• ナノディスク再構成型鉄還元膜タンパク質101F6における電子移動反応の解析

  山口葵, Abosharaf Hamed A., 鍔木基成, 木村哲就

  第94回日本生化学大会, Nov. 2021, Japanese, 日本生化学会, Online, Domestic conference, Co-authored internationally

  Poster presentation


• Double spin-label ESR spectroscopic analysis of structural changes in heme ABC transporter induced by nucleotide-binding.

  仲絢香, 小堀康博, 鍔木基成, 城宜嗣, 杉本宏, 木村哲就

  第94回日本生化学大会, Nov. 2021, Japanese, 日本生化学会, Online, Domestic conference

  Poster presentation


• 蛍光寿命測定によるα-Synucleinの脂質二重膜結合状態の構造解析

  笹田航, 松原亮介, 藤井宏一, 木村哲就

  第21回日本蛋白質科学会年会, Jun. 2021, Japanese, 日本蛋白質科学会, Online, Domestic conference

  Poster presentation


• 二重スピンラベルESR分光法を用いたヘムインポーターBhuUV-Tの構造ダイナミクスの解析

  仲絢香, 小堀康博, 鍔木基成, 城宜嗣, 杉本宏, 木村哲就

  第21回日本蛋白質科学会年会, Jun. 2021, Japanese, 日本蛋白質科学会, Online, Domestic conference

  Poster presentation


• 101F6 ナノディスクを用いた電子移動反応の実験的解析

  山口葵, Abosharaf Hamed A., 鍔木基成, 木村哲就

  第21回日本蛋白質科学会年会, Jun. 2021, Japanese, 日本蛋白質科学会, Online, Domestic conference, Co-authored internationally

  Poster presentation


• ナノディスク再構成型BhuUV-Tによる段階的な基質輸送の分光学的観察

  木村哲就, 浅田拓也, 林沙英, 鍔木基成, 城宜嗣, 杉本宏

  第21回日本蛋白質科学会年会, Jun. 2021, Japanese, 日本蛋白質科学会, Online, Domestic conference

  Poster presentation


• ナノディスク再構成型BhuUV-Tによる段階的な基質輸送の分光学的観察

  木村哲就, 浅田拓也, 林沙英, 鍔木基成, 城宜嗣, 杉本宏

  第21回日本蛋白質科学会年会, Jun. 2021, Japanese, 日本蛋白質科学会, Online, Domestic conference

  Oral presentation


• ヘムABCトランスポーターBhuUVの輸送機構に関する分光学的解析

  木村哲就, 浅田拓也, 仲絢香, 林沙英, 城宜嗣, 杉本宏

  第47回生体分子科学討論会, Jun. 2021, Japanese, 第47回生体分子科学討論会実行委員会, Online, Domestic conference

  Oral presentation


• 101F6ナノディスクを用いた電子移動反応の実験的解析

  山口葵, Abosharaf Hamed A., 鍔木基成, 木村哲就

  第47回生体分子科学討論会, Jun. 2021, Japanese, 第47回生体分子科学討論会実行委員会, Online, Domestic conference, Co-authored internationally

  Poster presentation


• 二重スピンラベルESR分光法を用いたABCトランスポーターBhuUVの構造解析

  仲絢香, 小堀康博, 鍔木基成, 城宜嗣, 杉本宏, 木村哲就

  第47回生体分子科学討論会, Jun. 2021, Japanese, 第47回生体分子科学討論会実行委員会, Online, Domestic conference

  Poster presentation


• α-Synucleinの脂質二重膜結合状態の分光学的構造解析

  笹田航, 松原亮介, 藤井宏一, 木村哲就

  第47回生体分子科学討論会, Jun. 2021, Japanese, 第47回生体分子科学討論会実行委員会, Online, Domestic conference

  Poster presentation


• 分子拡散混合を利用したヘムABCトランスポーターの時間分解分光解析

  KIMURA Tetsunari

  ピコバイオロジー研究会, Dec. 2020, Japanese, 兵庫県立大ピコバイオロジー研究所, Kamigori, Domestic conference

  [Invited]

  Invited oral presentation


• Structural and functional dynamics of ABC transporters clarified by novel time-resolved measurement systems

  KIMURA Tetsunari

  International Conference on Chemical and Environmental Sciences 2020, Dec. 2020, English, Institute of Engineering & Management, Kolkata, Online, International conference

  [Invited]

  Keynote oral presentation


• Time-resolved spectroscopic measurements on the transport dynamics of ABC transporter

  KIMURA Tetsunari, HAYASHI Sae, IKEMOTO Yuka, SHIRO Yoshitsugu, SUGIMOTO Hiroshi

  第58回日本生物物理学会年会, Sep. 2020, English, The Biophysical Soceity of Japan, Online, Domestic conference

  Poster presentation


• Enhancement of ferric reductase activity of human Steap3 upon reconstitution into phospholipid bilayer nanodisc

  NISHI Ayane, NAKATA SOUTO, TAKEUCHI Fusako, KIMURA Tetsunari, TSUBAKI Motonari

  第58回日本生物物理学会年会, Sep. 2020, English, The Biophysical Soceity of Japan, Online, Domestic conference

  Poster presentation


• Characterization of reaction intermediate formed in the microsecond time domain of the catalytic reaction of nitric oxide reductase

  TOSHA Takehiko, TAKEDA Hanae, KIMURA Tetsunari, HORITANI Masaki, SHIRO Yoshitsugu

  第58回日本生物物理学会年会, Sep. 2020, English, The Biophysical Soceity of Japan, Online, Domestic conference

  Poster presentation


• Spectroscopic analysis of allocrite transport mechanism using nanodisc-reconstituted heme ABC transporter

  ASADA Takuya, TSUBAKI Motonari, SHIRO Yoshitsugu, SUGIMOTO Hiroshi, KIMURA Tetsunari

  第58回日本生物物理学会年会, Sep. 2020, English, The Biophysical Soceity of Japan, Online, Domestic conference

  Poster presentation


• Structural changes of a-synuclein along the lipid-binding and oligomerization revealed by fluorescence lifetime measurements.

  SASADA Ko, FUJII Koichi, MATSUBARA Ryosuke, KIMURA Tetsunari

  第58回日本生物物理学会年会, Sep. 2020, English, The Biophysical Soceity of Japan, Online, Domestic conference

  Poster presentation


• Could the biogenic zinc oxide nanoparticles inhibit the ATPase activity of ABC transporters?

  Aliaa M. Radwan, Mai M. El-Keiy, Tarek M. Mohamed, 木村 哲就

  第58回日本生物物理学会年会, Sep. 2020, English, The Biophysical Soceity of Japan, Online, Domestic conference, Co-authored internationally

  Poster presentation


• Reconistitution of Cecytb-2 in Phospholipid Bilayer Nanodisc and Meaurments of its Ferric Reductase Activity

  ABOSHARAF Hamed A., SAKAMOTO Yuki, El BEHERY Mohammed, DIAB Thoria, MOHAMED Tarek M., KIMURA Tetsunari, TSUBAKI Motonari

  第58回日本生物物理学会年会, Sep. 2020, English, The Biophysical Soceity of Japan, Online, Domestic conference, Co-authored internationally

  Poster presentation


• 高速分子動画撮影を補完する時間分解分光測定法の開発と微結晶タンパク質試料への応用

  KIMURA Tetsunari

  第93回生化学会大会, Sep. 2020, Japanese, Japan Biochemistry Society, Online, Domestic conference

  [Invited]

  Invited oral presentation


• Observation of Structural Change Of α-Synuclein, an Intrinsically Disordered Protein by Fluorescence Lifetime Measurement

  SASADA Ko, FUJII Koichi, MATSUBARA Ryosuke, KIMURA Tetsunari

  第20回日本蛋白質科学会, Jul. 2020, English, Protein Science Society of Japan, 誌上, Domestic conference

  Poster presentation


• Membrane Protein Dynamics Studied by Time-resolved Measurements

  Tetsunari Kimura

  Indo-Japan Joint Workshop “Frontiers in Molecular Spectroscopy: From Fundamentals to Applications in Chemistry and Biology”, Nov. 2019, English, Kobe, International conference

  [Invited]

  Invited oral presentation


• Complex Formation and Heme Transport of BhuUV-T Investigated by Time-Resolved UV/vis Spectroscopy

  Tetsunari Kimura

  2019 Summer Progress Report Meeting Cellular Regulation Laboratory, Univ. Hyogo, Aug. 2019, English, Univ. Hyogo, Domestic conference

  Oral presentation


• 蛍光寿命測定によるヘリックスバンドルタンパク質ACBP の構造不均一性

  藤井 宏一, MATSUBARA RYOSUKE, TSUBAKI MOTONARI, KIMURA TETSUNARI

  第19回日本蛋白質科学会年会･第71回日本細胞生物学会大会 合同年次大会, Jun. 2019, Japanese, 神戸国際会議場, Domestic conference

  Poster presentation


• ナノディスク再構成型ヘムABC トランスポーターを用いた基質輸送機構の分光学的解析

  浅田 拓也, TSUBAKI MOTONARI, 城 宜嗣, 杉本 宏, KIMURA TETSUNARI

  第19回日本蛋白質科学会年会･第71回日本細胞生物学会大会 合同年次大会, Jun. 2019, Japanese, 神戸国際会議場, Domestic conference

  Poster presentation


• Elucidation of molecular functions of human tumor suppressor protein 101F6 by reconstitution into phospholipid bilayer nanodiscs

  Mohamed El-Behery, Akikazu Asada, Fusako Takeuchi, Tetsunari Kimura, Eri Chatani, ○Motonari Tsubaki

  34TH PHILIPPINE CHEMISTRY CONGRESS, May 2019, English, Waterfront Hotel and Convention Center（Philippines）, International conference

  Oral presentation


• タンパク質の化学的ダイナミズム

  KIMURA TETSUNARI

  第9回K-CONNEX研究会, Mar. 2019, Japanese, 神戸大学六甲台第1キャンパスアカデミア館5階504 特別演習室, Domestic conference

  [Invited]

  Invited oral presentation


• SPring-8における時間分解分光計測系の構築とタンパク質の分子機構解析への応用

  KIMURA TETSUNARI

  第38回SPring-8先端利用技術ワークショップ〜SPring-8放射光実験の生物・材料・無機化学への応用〜, Feb. 2019, Japanese, 神戸大学 六甲台キャンパス 自然科学総合研究棟1号館, Domestic conference

  [Invited]

  Invited oral presentation


• 部位特異的スピンラベルEPR分光法によるABCトランスポーター; BhuUVの構造変化測定

  大西 萌, KOBORI YASUHIRO, 城 宜嗣, 杉本 宏, TSUBAKI MOTONARI, KIMURA TETSUNARI

  神戸大学先端融合研究環境環ワークショップ『非共有結合系の分子科学：計測技術から探る生体分子科学の新展開』, Jan. 2019, Japanese, 神戸大学 六甲台キャンパス 理学部Z201, 205号室, Domestic conference

  Poster presentation


• 蛍光寿命計測によるACBPのフォールディング機構解析

  藤井 宏一, MATSUBARA RYOSUKE, TSUBAKI MOTONARI, KIMURA TETSUNARI

  神戸大学先端融合研究環境環ワークショップ『非共有結合系の分子科学：計測技術から探る生体分子科学の新展開』, Jan. 2019, Japanese, 神戸大学 六甲台キャンパス 理学部Z201, 204号室, Domestic conference

  Poster presentation


• マイクロ流路ミキサーを利用したタンパク質の実時間観察

  KIMURA TETSUNARI

  神戸大学先端融合研究環境環ワークショップ『非共有結合系の分子科学：計測技術から探る生体分子科学の新展開』, Jan. 2019, Japanese, 神戸大学 六甲台キャンパス 理学部Z201, 202号室, Domestic conference

  [Invited]

  Invited oral presentation


• Spectroscopic studies on the transport mechanism of heme ABC transporter

  林 沙英, 城 宜嗣, 杉本 宏, TSUBAKI MOTONARI, KIMURA TETSUNARI

  神戸大学先端融合研究環境環ワークショップ『非共有結合系の分子科学：計測技術から探る生体分子科学の新展開』, Jan. 2019, Japanese, 神戸大学 六甲台キャンパス 理学部Z201, 203号室, Domestic conference

  Poster presentation


• 部位特異的スピンラベルEPR分光法によるABCトランスポーター;BhuUVの構造変化の実時間測定

  大西 萌, TSUBAKI MOTONARI, KOBORI YASUHIRO, KIMURA TETSUNARI

  神戸大学 研究基盤センター 若手フロンティア研究会2018, Dec. 2018, Japanese, 神戸大学百年記念館, Domestic conference

  Poster presentation


• 蛍光寿命計測によるアシルCoA結合タンパク質のフォールディング機構解析

  藤井 宏一, MATSUBARA RYOSUKE, TSUBAKI MOTONARI, KIMURA TETSUNARI

  若手フロンティア研究会2018, Dec. 2018, Japanese, 神戸大学百年記念館, Domestic conference

  Poster presentation


• 分光法による膜タンパク質のダイナミクス観察

  KIMURA TETSUNARI

  平成30 年度膜タンパク質研究会, Oct. 2018, Japanese, 淡路夢舞台国際会議場, Domestic conference

  [Invited]

  Invited oral presentation


• Membrane proteins in action; Structural and functional dynamics revealed by time-resolved measurements.

  KIMURA TETSUNARI

  International Congress on Pure and Applied Chemistry 2018, Oct. 2018, English, Langkawi, Malaysia, International conference

  Keynote oral presentation


• 線虫Cytochrome b561ホモログCecytb-2のアスコルビン酸由来電子伝達反応解析

  福澤 美咲, 藤村 美香, 三浦 雅央, KIMURA TETSUNARI, TSUBAKI MOTONARI

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

  Poster presentation


• Vibrational analysis for studying ion-protein interactions of a magnesium ion channel, MgtE

  Tetsunari Kimura, Victor Lorenz-Fonfria, Shintaro Doki, Hideyoshi Motoki, Ryuichiro Ishitani, Osamu Nureki, Masahiro Higashi, ○Yuji Furutan

  第56回日本生物物理学会年会, Sep. 2018, English, 岡山大学 津島キャンパス, Domestic conference

  Poster presentation


• Real-time measurements of the conformational changes in ABC transporter; BhuUV, revealed by site-directed spin-labeling EPR spectroscopy

  ○Kizashi Onishi, Motonari Tsubaki, Yasuhiro Kobori, Tetsunari Kimura

  第56回日本生物物理学会年会, Sep. 2018, English, 岡山大学 津島キャンパス, Domestic conference

  Poster presentation


• Folding dynamics of acyl-CoA binding protein revealed by fluorescence lifetime measurements

  ○Koichi Fujii, Motonari Tsubaki, Tetsunari Kimura

  第56回日本生物物理学会年会, Sep. 2018, English, 岡山大学 津島キャンパス, Domestic conference

  Poster presentation


• Direct observation of the enzymatic reaction catalyzed by an integral membrane NO reductase using microflow-flash IR spectroscopy

  Hanae Takeda, Tetsunari Kimura, Takashi Nomura, Shoko Ishii, Akiko Matsubayashi, Azusa Yokota, Takehiko Tosha, Yoshitsugu Shiro, Minoru Kubo

  第56回日本生物物理学会年会, Sep. 2018, English, 岡山大学 津島キャンパス, Domestic conference

  Oral presentation


• Cytochrome b561ホモログ・線虫Cecytb-2の三価鉄還元酵素活性の分子機構

  藤村 美香, 三浦 雅央, KIMURA TETSUNARI, TSUBAKI MOTONARI

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

  Poster presentation


• Analyses on the ascorbate-specific electron transfer function of Cecytb-2, a cytochrome b561 homolog in Caenorhabditis elegans

  ○Misaki Fukuzawa, Mika Fujimura, Masahiro Miura, Tetsunari Kimura, Motonari Tsubaki

  第56回日本生物物理学会年会, Sep. 2018, English, 岡山大学 津島キャンパス, Domestic conference

  Poster presentation


• 線虫Cytohrome b561ホモログCecytb-2分子機能解析

  Misaki Fukuzawa, Mika Fujimra, Masahiro Miura, Tetsunari Kimura, TSUBAKI MOTONARI

  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、神戸大学百年記念館, Domestic conference

  Poster presentation


• 線虫Cytochrome b561ホモログCecytb-2のアスコルビン酸からの電子伝達機能解析

  福澤 美咲, 藤村 美香, 三浦 雅央, TSUBAKI MOTONARI, KIMURA TETSUNARI

  2017年度生命科学系学会合同年次大会, Dec. 2017, Japanese, 神戸ポートアイランド, Domestic conference

  Poster presentation


• ヘムABCトランスポーターの輸送機構に関する分光学的研究

  林 沙英, KIMURA TETSUNARI

  2017年度生命科学系学会合同年次大会, Dec. 2017, Japanese, 神戸ポートアイランド, Domestic conference

  Poster presentation


• Cytochrome b561ホモログ・線虫Cecytb-2の分子機能解明

  藤村 美香, 三浦 雅央, KIMURA TETSUNARI, TSUBAKI MOTONARI

  2017年度生命科学系学会合同年次大会, Dec. 2017, Japanese, 神戸ポートアイランド, Domestic conference

  Poster presentation


• ABCトランスポーターのヘム輸送機構解析

  Sae Hayashi, Hiroshi Sugimoto, Yoshitsug Shiro, Yuka Ikemoto, TSUBAKI MOTONARI, Tetsunari Kimura

  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、神戸大学百年記念館, Domestic conference

  Poster presentation


• Elucidation of the molecular function of Caenorhabditis elegans Cecytb-2, a cytochrome b561 homologue

  Mika Fujimura, Masahiro Miura, KIMURA TETSUNARI, TSUBAKI MOTONARI

  第55回日本生物物理学会年会, Sep. 2017, Japanese, 熊本大学 黒髪北地区, Domestic conference

  Poster presentation


• Direct observation of allocate-transport and ATP-hydrolysis for the ABC transport by time-resolved spectroscopy

  KIMURA TETSUNARI, HAYASHI SAE, SHIRO YOSHITSUGU, SUGIMOTO HIROSHI, IKEMOTO YUKA

  第55回日本生物物理学会年会, Sep. 2017, Japanese, 熊本大学 黒髪北地区, Domestic conference

  [Invited]

  Invited oral presentation


• Elucidation of the NO Reduction Mechanism of Nitric Oxide Reductase Using Time-resolved Vis / IR Spectroscopy

  Hanae Takeda, KIMURA TETSUNARI, Takashi Nomura, Akiko Matsubayashi, Shoko Ishii, Takehiko Tosha, Yoshitsugu Shiro, Minoru Kubo

  第55回日本生物物理学会年会, Sep. 2017, Japanese, 熊本大学 黒髪北地区, Domestic conference

  Poster presentation


• 線虫cytochrome b561ホモログCecytb-2の分子機能解明

  FUJIMURA MIKA, MIURA MASAHIRO, KIMURA TETSUNARI, TSUBAKI MOTONARI

  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, 神戸大学百年記念館、神戸、兵庫県, Domestic conference

  Poster presentation


• Intermediate structures of the active site in membrane proteins revealed by time-resolved spectroscopy with micro-channel devices.

  Tetsunari Kimura

  シンポジウム:時空間精密構造解析による生体分子活性サイトの機能解明(第54回日本生物物理学会年会), Nov. 2016, English, つくば国際会議場, Domestic conference

  Oral presentation


• Detection of Short-Lived Reaction Species of Nitric Oxide Reductase Using Nanoliter-Flow Time-Resolved Visible/IR Spectroscopy

  Hanae Takeda, Tetsunari Kimura, Shoko Ishii, Takehiko Tosha, Yoshitsugu Shiro, Minoru Kubo

  第54回日本生物物理学会年会, Nov. 2016, English, つくば国際会議場, Domestic conference

  Poster presentation


• Cytochrome b561ホモログ・線虫Cecytb-2の分子機能解明

  FUJIMURA MIKA, MIURA MASAHIRO, KIMURA TETSUNARI, TSUBAKI MOTONARI

  第89回日本生化学会大会, Sep. 2016, Japanese, 宮城県、仙台国際センター、東北大学, Domestic conference

  Poster presentation


• マイクロ流路フロー・フラッシュ時間分解赤外分光法による一酸化窒素還元酵素の反応機構解析

  KIMURA TETSUNARI, 武田英恵, 石井頌子, 當舎武彦, 城宜嗣, 久保稔

  生体分子科学討論会, Jun. 2016, Japanese, 名古屋大学 野依記念学術交流館, Domestic conference

  Oral presentation


• NO reduction mechanism in the binuclear catalytic center of nitric oxide reductase revealed by novel micro-channel flow-flash spectrometers

  Tetsunari KIMURA, Hanae TAKEDA, Shoko ISHII, Takehiko TOSHA, Yoshitsugu SHIRO, Minoru KUBO

  Metallocofactors Gordon Research Conference, Jun. 2016, English, Stonehill College Easton, MA, International conference

  Poster presentation


• Molecular mechanism of the catalytic reaction of NO reductase revealed by novel time-resolved visible/IR absorption spectrometers with microfluidic device.

  KIMURA Tetsunari, TAKEDA Hanae, ISHII Shoko, TOSHA Takehiko, SHIRO Yoshitsugu, KUBO Minoru

  Biophysical Soceity 60th Annual Meeting, Feb. 2016, English, Biophysical Society, Los Angeles, CA, USA, Time-resolved (TR) spectroscopy plays convincing roles in clarifying the molecular mechanism of biological reactions in the atomic and electronic level. Most of the biological reactions can be triggered by the sudden changes in buffer conditions, but the time-resolution of the conventional solution-mixing technique is limited to several milliseconds and the sample consumption i, International conference

  Poster presentation


• Stopped-flow kinetic analysis of human duodenal cytochrome b561

  TAKEDA Hanae, TOGASHI Hiromi, KIMURA Tetsunari, Grant Mauk, SUGIMOTO Hiroshi, 城 宜嗣SHIRO Yoshitsugu

  Pacifichem2015 (The International Chemical Congress of Pacific Basin Societies 2015), Dec. 2015, English, American Chemical Society, Chemical Society of Japan, 他5カ国の化学会, Honolulu, HI, USA, International conference

  Poster presentation


• Molecular mechanism of NO reduction by bacterial nitric oxide reductases

  SHIRO Yoshitsugu, TOSHA Takehiko, KIMURA Tetsunari, KUBO Minoru

  Pacifichem2015 (The International Chemical Congress of Pacific Basin Societies 2015), Dec. 2015, English, American Chemical Society, Chemical Society of Japan, 他5カ国の化学会, Honolulu, HI, USA, International conference

  [Invited]

  Invited oral presentation


• Development of a time-resolved visible/IR absorption spectrometer with a novel microfluidic device and its application to catalytic reaction of NO reductase.

  KIMURA Tetsunari, ISHII Shoko, TOSHA Takehiko, SHIRO Yoshitsugu, KUBO Minoru

  Pacifichem2015 (The International Chemical Congress of Pacific Basin Societies 2015), Dec. 2015, English, American Chemical Society, Chemical Society of Japan, 他5カ国の化学会, Honolulu, HI, USA, Time-resolved (TR) spectroscopy plays convincing roles in clarifying the molecular mechanism of biological processes in molecular and atomic level, and it has been applied to observe various photocyclic reactions with the laser flash technique. However, most of the biological reactions are non-photocyclic and irreversible, and the applications of TR spectroscopy to such reactio, International conference

  Poster presentation


• Development of Micro-channel Flow-flash Infrared Absorption Spectroscopy and its Application to Intermediate of Nitric Oxide Reductase.

  KIMURA Tetsunari, ISHII Shoko, TOSHA Takehiko, SHIRO Yoshitsugu, KUBO Minoru

  日本生物物理学会第52回年会, Sep. 2015, English, The Biophysical Soceity of Japan, Kanazawa University, Characterization of the transient intermediates in the chemical and biological reactions is essential to clarify their molecular mechanisms. In this research, we have developed an infrared spectroscopic system by combining laser spectroscopy and micro-channel flow-cell, which enabled us to monitor the irreversible enzymatic reaction of low-yield membrane protein with the nanose, Domestic conference

  Oral presentation


• マイクロ流体フロー・フラッシュ赤外分光測定系による一酸化窒素還元酵素の触媒反応の実時間観察

  KIMURA Tetsunari, ISHII Shoko, TOSHA Takehiko, SHIRO Yoshitsugu, KUBO Minoru

  第9回分子科学討論会, Sep. 2015, Japanese, Japan Society for Molecular Science, Tokyo Institute of Technology, 酵素反応の分子機構解明には、反応過程の実時間観察が重要な役割を果たす。しかし、従来の装置では基質の結合から始まる酵素反応の動的過程においてミリ秒以内に生成する反応初期の中間体を観測することは困難である。そこで、マイクロ流体デバイスと赤外レーザーを組み合わせたフロー・フラッシュ赤外分光測定系を開発し、マイクロ秒の時間分解能での観察を可能にした。本研究では、装置を一酸化窒素還元酵素(NOR)に応用し、ヘム鉄・鉄原子から構成される活性中心においてNOがN2Oへと還元される触媒反応過程を、基質のNO伸縮振動・NN伸縮振動に着目して追跡した。当日は、速度論的解析と併せ、NORの反応機構について議論する。, Domestic conference

  Oral presentation


• マイクロ流体フローを用いた一酸化窒素還元酵素の触媒反応の動的分光観察 (招待講演)

  KIMURA Tetsunari

  第15回日本蛋白質科学会年会, Jun. 2015, Japanese, Protein Science Society of Japan, Tokushima, 一酸化窒素還元酵素は、ヘム鉄と非ヘム鉄からなる活性中心で2分子のNOをN2Oに還元する膜蛋白質であり、細胞毒性の高いNOを速やかに無毒化する。近年、我々は結晶構造解析から、本酵素の活性中心の構造や電子・プロトン供給経路を明らかにした。一方、反応機構に関しては、これまでに、(i) 2分子のNOがヘム鉄と非ヘム鉄それぞれに結合した後、N-N間に共有結合が形成されるtrans機構と、(ii) 1分子のNOがヘム鉄と非ヘム鉄を架橋するように結合した後、2分子目のNOがNOを攻撃するcis機構が提案されており、まだ議論がなされている。これは機構解明の鍵となる過渡的中間体; NO結合型が1 ms以内に生成・消失し、従来の方法では構造解析が不可能なためである。そこで、マイクロ秒の時間分解能での計測が可能な顕微可視・赤外吸収分光装置を新規に開発し、中間体を直接観察す, Domestic conference

  [Invited]

  Invited oral presentation


• マイクロ流体フローを用いた一酸化窒素還元酵素の触媒反応の動的分光観察 (ポスター)

  KIMURA Tetsunari

  第15回日本蛋白質科学会年会, Jun. 2015, Japanese, Protein Science Society of Japan, Tokushima, 一酸化窒素還元酵素は、ヘム鉄と非ヘム鉄からなる活性中心で2分子のNOをN2Oに還元する膜蛋白質であり、細胞毒性の高いNOを速やかに無毒化する。近年、我々は結晶構造解析から、本酵素の活性中心の構造や電子・プロトン供給経路を明らかにした。一方、反応機構に関しては、これまでに、(i) 2分子のNOがヘム鉄と非ヘム鉄それぞれに結合した後、N-N間に共有結合が形成されるtrans機構と、(ii) 1分子のNOがヘム鉄と非ヘム鉄を架橋するように結合した後、2分子目のNOがNOを攻撃するcis機構が提案されており、まだ議論がなされている。これは機構解明の鍵となる過渡的中間体; NO結合型が1 ms以内に生成・消失し、従来の方法では構造解析が不可能なためである。そこで、マイクロ秒の時間分解能での計測が可能な顕微可視・赤外吸収分光装置を新規に開発し、中間体を直接観察す, Domestic conference

  Poster presentation


• Stopped-Flow Analysis on the Reaction of Ascorbate with Duonal Cytochrome b561

  TAKEDA Hanae, TOGASHI Hiromi, KIMURA Tetsunari, Grant Mauk, 城 宜嗣SHIRO Yoshitsugu, SUGIMOTO Hiroshi

  第15回日本蛋白質科学会年会, Jun. 2015, Japanese, Protein Science Society of Japan, Tokushima, Domestic conference

  Poster presentation


• Preparation of Fungal NO-Reductase Crystals in the Intermediate State Using Caged-NO

  NISHIDA Takuma, TOSHA Takehiko, SAKAGUCHI Miyuki, KIMURA Tetsunari, YANAGISAWA Sachiko, UENO Tsuyoshi, MURAKAMI Hironori, YAMAMOTO Masaki, OGURA Takashi, SHIRO Yoshitsugu, KUBO Minoru

  第15回日本蛋白質科学会年会, Jun. 2015, Japanese, Protein Science Society of Japan, Tokushima, Domestic conference

  Poster presentation

• Protein Science Society of Japan

  2003 - Present


• 生物物理学会（Biophysical Society, U.S.A.）

  2002 - Present


• 日本生物物理学会

  2000 - Present


• 日本化学会

• 新しい動的構造解析法の開発とABCトランスポーターへの応用

  杉本 宏, 木村 哲就

  日本学術振興会, 科学研究費助成事業, 基盤研究(B), 国立研究開発法人理化学研究所, 01 Apr. 2024 - 31 Mar. 2027


• Development of Single-Molecule Reaction Imaging Techniques for Visualizing Carbon Resource Transformation

  立川 貴士, 隈部 佳孝, 松原 亮介, 木村 哲就

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research, Fund for the Promotion of Joint International Research (International Collaborative Research), Kobe University, 08 Sep. 2023 - 31 Mar. 2027


• Time-resolved measurements of allocrite transport in ABC transporters

  木村 哲就

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research, Grant-in-Aid for Scientific Research (B), Kobe University, 01 Apr. 2022 - 31 Mar. 2025


• 病原菌ヘムトランスポーターの構造変化の追跡と基質輸送機構の解明

  杉本 宏, 木村 哲就

  日本学術振興会, 科学研究費助成事業 基盤研究(B), 基盤研究(B), 国立研究開発法人理化学研究所, 01 Apr. 2021 - 31 Mar. 2024

  病原菌は宿主に感染した際に、血液ヘモグロビンから鉄をヘムの形で奪い取り、増殖に必要な栄養素として利用している。そのため、ヘムの取り込みは感染症を抑制する上でも注目されてきた。本課題では、グラム陰性の病原性細菌でアデノシン三リン酸（ATP）依存的にペリプラズム空間から細胞質へとヘムの膜輸送を行うABCトランスポーターの構造変化に着目する。これまでの当グループによる研究で明らかにした結晶構造からは、分子の中心を通る基質輸送チャネルに大きな構造変化が起こることが示唆されたが、ヘムやATPが結合した状態での構造は未だに決定できていないせいもあり、メカニズムの理解が全く進んでいない。本年度は、ヘムトランスポーターによるヘム輸送の仕組みの全容を解明するために、クライオ電子顕微鏡によって、ヘムの輸送過程の各構造のスナップショットを取得することを目指した。休止型の状態のトランスポーター（ペリプラズムヘム結合タンパク質：膜貫通サブユニット：ヌクレオチド結合サブユニットの1:2:2複合体）は比較的安定した状態であるため、試料調整方法の確立とデータ収集をおこなった。ヘム結合型の複合体の状態で界面活性剤を両親媒性ポリマーに置換することで可溶化状態を維持すると観察グリッド上での単分散性が向上することを見出した。凍結試料のクライオ電子顕微鏡観察を行なった結果、低分解能ではあるが5つのサブユニットの配置や２次構造が確認できた。


• 時間分解構造解析を補完する精密顕微分光計測

  久保 稔, 木村 哲就, 古谷 祐詞

  日本学術振興会, 科学研究費助成事業 新学術領域研究(研究領域提案型), 新学術領域研究(研究領域提案型), 兵庫県立大学, 28 Jun. 2019 - 31 Mar. 2024

  本研究では溶液測定に加えて、タンパク質微結晶を測定できる顕微分光技術を開発し、時分割結晶構造解析（分子動画法）と時分割分光法（紫外可視吸収分光、ラマン・赤外分光、蛍光分光）との相関的・補完的解析を推進する。多くの研究試料を扱うが、それらは新学術領域「高速分子動画」において研究されている試料である。以下、各分光法毎に本年度の装置開発実績及び研究実績を記す。
  ・紫外可視吸収分光：（微結晶測定）光遺伝学ツール・チャネルロドプシンの結晶相におけるダイナミクスを解明した。その分光データと時分割結晶構造解析との相関解析により、チャネルロドプシンの光誘起チャネル開口のメカニズムを解明した。新規ロドプシン類の測定も終えており、時分割結晶構造解析との相関解析を進めている。（溶液測定）顕微装置にマイクロ流路デバイスを組み込んだ極微量フロー測定により、嫌気呼吸の鍵酵素cNORの反応中間体同定に成功した。 ・赤外分光：（微結晶測定）微結晶に適用できる顕微時分割赤外分光装置の開発を完了した。N2O発生酵素P450norに装置を適用し、反応中間体の電子状態解明に成功した。（溶液測定）DNAフォトリアーゼの反応中間体を捉えることに成功した。現在、反応中間体の化学構造決定を目指し、同位体試料の測定を試みている。 ・ラマン分光：（微結晶測定）昨年度に引き続き、時分割顕微測定系の開発を進めている。一方、凍結結晶に対する測定系を整備し、P450酵素への適用を進めている。（溶液測定）二液混合系のstopped-flow測定装置を完成させた。 ・蛍光分光：（微結晶測定）二液混合系の時分割蛍光分光装置に分光器を導入した。今後、Ca2+感受性イクオリンに適用する。（溶液測定）Stopped-flow法を用いて、イクオリンの蛍光ダイナミクスを測定した。現在、発光過程の分子モデルを構築するための速度論解析を進めている。


• タンパク質ダイナミクス観測に資する高時空間分解能ESR・IR計測系の開発と応用

  木村 哲就

  科学研究費補助金／新学術領域研究, Apr. 2017 - Mar. 2019, Principal investigator

  Competitive research funding


• ヒト癌抑制遺伝子候補101F6によるレドックス擾乱細胞死誘導機構の解明

  鍔木 基成

  学術研究助成基金助成金／基盤研究(C), Apr. 2016 - Mar. 2019

  Competitive research funding


• Development of a micro-flow time-resolved IR spectrometer for analyzing short-lived reaction intermediates of enzymes

  Kubo Minoru, TOSHA Takehiko, KIMURA Tetsunari

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B), Grant-in-Aid for Scientific Research (B), Institute of Physical and Chemical Research, 01 Apr. 2015 - 31 Mar. 2018

  This project aims to establish a time-resolved IR spectroscopic technique for studying dynamic structures of enzymes. A novel micro flow system has been developed and incorporated into a femtosecond laser-based IR spectrometer, which allows micro flow-flash IR measurements of enzymatic reactions with the minimum sample volume of 20 nL. We applied this measurement system to NO reductase, a heme enzyme that catalyzes the reduction of NO to N2O in the nitrogen cycle, and succeeded in determining the NO binding mode in the short-lived intermediate. Furthermore, we also used the micro-flow flash method for time-resolved crystallography at SACLA, and succeeded in visualizing the 3D structure of the NO-bound intermediate during the NO reduction reaction.


• 新規時間分解計測法の開発とABCトランスポーターの分子機構解析への応用

  木村 哲就

  科学研究費補助金／若手研究(A), Apr. 2015 - Mar. 2018, Principal investigator

  Competitive research funding


• Functional expression of mammalian ion channels and analysis of the molecular mechanisms

  FURUTANI Yuji, NAKAJO Koichi, TSUKAMOTO Hisao, KIMURA Tetsunari, KUBO Yoshihiro

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research, Grant-in-Aid for Challenging Exploratory Research, Institute for Molecular Science, 01 Apr. 2012 - 31 Mar. 2014

  Ion channel proteins work for selective permeation of ions in response to various stimuli and regulate electrical signals in living organisms. To understand the molecular mechanisms of these channels, the precise structural information with atomic resolution is required. In this study, mammalian ion channel proteins, which are important targets in medical and pharmaceutical sciences, have been studied by attenuated total reflection (ATR) FT-IR spectroscopy which gives us precise molecular information. After screening of several ion channel proteins, we have succeeded to resolve amide I bands of selectivity filter of KCNK1 channel, which is involved in regulation of resting membrane potential of cell, by using ATR FT-IR spectroscopy.


• Development of microfluidic mixer for time-resolved spectroscopy applicable to low-yield biomolecules

  KIMURA Tetsunari

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Young Scientists (A), Grant-in-Aid for Young Scientists (A), 01 Apr. 2011 - 31 Mar. 2014

  To understand the molecular mechanism of biomolecular reactions, the dynamics of the molecules along the reaction have to be revealed by the real-time measurements. However, the yield of protein is usually very low, ug-mg, and the time-resolved measurements is hard to apply for many biomolecular reactions because of the relatively large consumption. In this research, the time-resolved spectroscopies applicable to the low-yield protein samples were developed and applied to observe the folding, membrane transport and enzymatic reactions.


• 時間分解赤外分光法を用いた膜蛋白質フォールディング機構の解明

  木村 哲就

  日本学術振興会, 科学研究費助成事業 研究活動スタート支援, 研究活動スタート支援, 分子科学研究所, 2010 - 2011

  本申請研究は、光開裂・解離性分子をβバレル膜蛋白質OmpAに修飾することで蛋白質構造に大きな摂動を加え、それによって引き起こされた変性状態からのフォールディングを生理的条件に近い温度・pH・塩濃度環境下において追跡することを目的とした。そこで、まずOmpAのβバレル構造ドメインの遺伝子配列をpETベクターに組み込んだプラスミドを作成し、そのプラスミドを形質転換した大腸菌を用いることによって発現を試みた。OmpAのβバレルドメインの発現をSDS-PAGEによって確認した。さらに簡便な精製を目的としてHisタグをC末端側に導入したプラスミドを作成すると同時に、システイン残基を導入した変異体の作製も行った。今後、光解離性分子によるOmpAの修飾を行い、構造を詳細に検討する予定である。一方、フォールディング反応に伴う膜蛋白質の構造変化に関する詳細な情報を得るためには、時間分解赤外差スペクトル分光測定を駆使する予定である。さらには、フォールディングキネティクスの解析には、得られた一連の赤外差スペクトルに対してSVD解析とグローバルフィッティング法を用いて、中間体の構造も含めて検討する必要がある。そこで、αヘリックスから構成される膜蛋白質pHRに関して、その光反応サイクルを実時間観察し、詳細な解析を行うことによって、膜蛋白質構造において起こる変化を追跡することを試みた。その結果、溶媒から隔離されたヘリックスの構造変化が光反応サイクルの後期に起こることがわかった。この予備実験により、よりドラスティックな構造変化であるフォールディング反応を時間分解赤外差スペクトル分光測定によってとらえることができると期待される。


• マイクロ秒分割分光装置を用いた蛋白質の折り畳み機構の解明

  基礎科学特別研究員研究費, 2003 - 2005

  Competitive research funding


• 圧力効果を利用した蛋白質の折り畳み機構における水分子ダイナミクスの直接観察

  基礎科学特別研究員研究費, 2005

  Competitive research funding