Updated on 2026/03/10
膜タンパク質のX線結晶構造解析を大阪大学・蛋白質研究所・月原冨武の下で学ぶ。また、膜タンパク質の精製、結晶化は学生時代の大半を過ごした姫路工業大学（現・兵庫県立大学）・理学部・吉川信也の下で学ぶ。学位取得後、大阪大学・産業科学研究所・山口明人の主宰する研究室で多剤排出トランスポーターの構造研究に従事。世界初の多剤排出トランスポーターの構造を決定。以来、多剤排出トランスポーターなどの膜タンパク質を中心に構造機能研究を続けている。
バークホルデリア菌の多剤排出ポンプの結晶構造解析に成功 殺人バクテリアの薬剤耐性（AMR）対策に向けて
2023/07/11
Languages： Japanese
要点-殺人バクテリア（バークホルデリア菌）の薬剤耐性（AMR）機構の構造と働きを解明。-バークホルデリア菌AMRの主因のひとつである多剤排出ポンプの結晶構造解析に成功。-多剤排出ポンプの働きを抑える新薬や、排出されにくい抗生物質の開発に期待。概要
肺炎桿菌の多剤排出ポンプの結晶構造解析に成功 コロナ感染の二次感染菌に備わる多剤耐性化機構の解明
2021/09/29
Languages： Japanese
要点-肺炎桿菌の多剤排出ポンプの立体構造を原子レベルで解析することに成功。-多剤排出ポンプが薬剤を排出する仕組みの解明で、多剤耐性化の克服へ前進。-コロナ死者の大多数に認められる二次的な肺炎桿菌感染症に対する、有効な抗菌薬治療に期待。概要東京工業大学（以下「東工大」）生命理工学院生命理工学系の村上聡
多剤排出トランスポーターの薬剤排出機構を解明 スーパーコンピュータ「京」で巨大分子機械の動きを計算
2018/03/14
Languages： Japanese
理化学研究所（理研） 計算科学研究機構 粒子系生物物理研究チームの松永康佑研究員（JSTさきがけ研究員）、横浜市立大学 大学院の山根努特任助教、池口満徳教授、木寺詔紀教授、東京工業大学の村上聡教授らの共同研究グループ※は、社会問題となっている多剤耐性細菌の原因の1つとされる多剤排出トランスポーター「AcrB」の薬剤排出機構を、スーパーコンピュータ「京」を用いたシミュレーションによって解明しました。
マクロライド排出ポンプの結晶構造解析に成功―マクロライド系抗生物質の排出、病原性因子の分泌機構解明に光―
2017/11/10
Languages： Japanese
病原性細菌が持つ薬剤を排出するポンプの新しい構造が明らかになった マクロライド系薬剤排出の仕組みが分かることで、マクロライド耐性克服の可能性 病原性細菌が病原性の原因物質を分泌するためのポンプでもあり、その働きを抑制すれば病原菌の病原性を軽減できる可能性
Structural Biology
Protein Crystallography
生物物理学
Life Science / Biophysics
Life Science / Structural biochemistry
Osaka University
- 1997
Tokyo Institute of Technology Bioscience and Biotechnology Professor
2008
ケンブリッジ大学 客員研究員
2006 - 2010
Osaka University The Institute of Scientific and Industrial Research Associate Professor
2004 - 2008
科学技術振興機構 さきがけ研究者
2002 - 2006
Osaka University The Institute of Scientific and Industrial Research
1998 - 2004
Radha Nandishaiah, Satoshi Murakami, Shahul Hameed P, Maho Aoki, Ui Okada, Eiki Yamashita, Suryanarayanan Venkatesan, Nagakumar Bharatham, Sudipta Sarma, Anirudh P. Shanbhag, Sreevalli Sharma, Ranga Rao, Vasanthi Ramachandran, V. Balasubramanian, Santanu Datta, Nainesh Katagihallimath
Communications Biology 8 ( 1 ) 2025.11
Mariya Lazarova, Thomas Eicher, Clara Börnsen, Hui Zeng, Mohd Athar, Ui Okada, Eiki Yamashita, Inga M. Spannaus, Max Borgosch, Hi-jea Cha, Attilio V. Vargiu, Satoshi Murakami, Kay Diederichs, Achilleas S. Frangakis, Klaas M. Pos
2024.11
Crystal structure of topoisomerase IV from Klebsiella pneumoniae in complex with DNA and BWC0977, a dual-targeting broad-spectrum novel bacterial topoisomerase inhibitor.
S. Murakami, U. Okada, E. Yamashita, M. Aoki, S. Hameed, N. Katagihallimath, V. Balasubramanian, V. Ramachandran, S. Datta
Worldwide Protein Data Bank 2024.11
Crystal structure of mycolic acid transporter MmpL3 from Mycobacterium smegmatis complexed with indolcarboxamide
S. Murakami, U. Okada, E. Yamashita, M. Pieroni, E. Carosati
Worldwide Protein Data Bank 2024.10
RND型多剤排出トランスポーターの構造機能研究～輸送素過程の考察 Reviewed
村上 聡, 岡田有意, 山下栄樹
生物物理 64 ( 4 ) 185 - 189 2024.7
Crystal structure of multidrug efflux transporter OqxB from Klebsiella pneumoniae
S. Murakami, E. Yamashita, U. Okada
Worldwide Protein Data Bank 2024.6
Crystal structure of aminoglycoside efflux transporter MexY from Pseudomonas aeruginosa
S. Murakami, E. Yamashita, U. Okada, M. Aoki
Worldwide Protein Data Bank 2024.4
Crystal Structure of the Multidrug effulx transporter BpeF from Burkholderia pseudomallei.
T. Kato, L.-W. Hung, E. Yamashita, U. Okada, T.C. Terwilliger, S. Murakami
2023.7
Crystal structures of multidrug efflux transporters from Burkholderia pseudomallei suggest details of transport mechanism Reviewed
Takaaki Kato, Ui Okada, Li-Wei Hung, Eiki Yamashita, Heung-Bok Kim, Chang-Yub Kim, Thomas C. Terwilliger, Herbert P. Schweizer, Satoshi Murakami
Proceedings of the National Academy of Sciences 120 ( 29 ) 2023.7
Structure-Based Analysis of Transient Interactions between Ketosynthase-like Decarboxylase and Acyl Carrier Protein in a Loading Module of Modular Polyketide Synthase. International journal
Taichi Chisuga, Satoshi Murakami, Akimasa Miyanaga, Fumitaka Kudo, Tadashi Eguchi
ACS chemical biology 18 ( 6 ) 1398 - 1404 2023.5
Structural and functional characteristics of the tripartite ABC transporter Reviewed
Ui Okada, Satoshi Murakami
Microbiology 168 ( 11 ) 2022.11
Real-time monitoring of polyacrylamide gel electrophoresis by the shadowgraph technique. International journal
Satoshi Murakami, Hiroaki Adachi, Hiroyoshi Matsumura, Kazufumi Takano, Tsuyoshi Inoue, Yusuke Mori
Journal of biochemistry 2022.11
Structure and function relationship of OqxB efflux pump from Klebsiella pneumoniae. International journal
Nagakumar Bharatham, Purnendu Bhowmik, Maho Aoki, Ui Okada, Sreevalli Sharma, Eiki Yamashita, Anirudh P Shanbhag, Sreenath Rajagopal, Teby Thomas, Maitrayee Sarma, Riya Narjari, Savitha Nagaraj, Vasanthi Ramachandran, Nainesh Katagihallimath, Santanu Datta, Satoshi Murakami
Nature communications 12 ( 1 ) 5400 - 5400 2021.9
Structural and functional diversity calls for a new classification of ABC transporters. International journal
Christoph Thomas, Stephen G Aller, Konstantinos Beis, Elisabeth P Carpenter, Geoffrey Chang, Lei Chen, Elie Dassa, Michael Dean, Franck Duong Van Hoa, Damian Ekiert, Robert Ford, Rachelle Gaudet, Xin Gong, I Barry Holland, Yihua Huang, Daniel K Kahne, Hiroaki Kato, Vassilis Koronakis, Christopher M Koth, Youngsook Lee, Oded Lewinson, Roland Lill, Enrico Martinoia, Satoshi Murakami, Heather W Pinkett, Bert Poolman, Daniel Rosenbaum, Balazs Sarkadi, Lutz Schmitt, Erwin Schneider, Yigong Shi, Show-Ling Shyng, Dirk J Slotboom, Emad Tajkhorshid, D Peter Tieleman, Kazumitsu Ueda, András Váradi, Po-Chao Wen, Nieng Yan, Peng Zhang, Hongjin Zheng, Jochen Zimmer, Robert Tampé
FEBS letters 594 ( 23 ) 3767 - 3775 2020.12
Tripartite transporters as mechanotransmitters in periplasmic alternating‐access mechanisms Reviewed
Satoshi Murakami, Ui Okada, Hendrik W. Veen
FEBS Letters 2020.10
三者複合体形成型ABCトランスポーターMacBの構造解析 Reviewed
岡田 有意, 村上 聡
ファルマシア 56 ( 6 ) 499 - 503 2020
Crystallization of aspirin form II by femtosecond laser irradiation Reviewed
Yuka Tsuri, Mihoko Maruyama, Riki Fujimoto, Shino Okada, Hiroaki Adachi, Hiroshi Y. Yoshikawa, Kazufumi Takano, Satoshi Murakami, Hiroyoshi Matsumura, Tsuyoshi Inoue, Katsuo Tsukamoto, Masayuki Imanishi, Masashi Yoshimura, Yusuke Mori
Applied Physics Express 12 ( 1 ) 015507 - 015507 2019.1
Growth of high-quality metastable crystal of acetaminophen using solution-mediated phase transformation at low supersaturation Reviewed
Fujimoto Riki, Maruyama Mihoko, Mori Yoichiro, Okada Shino, Adachi Hiroaki, Yoshikawa Hiroshi Y, Takano Kazufumi, Murakami Satoshi, Matsumura Hiroyoshi, Inoue Tsuyoshi, Imanishi Masayuki, Tsukamoto Katsuo, Yoshimura Masashi, Mori Yusuke
JOURNAL OF CRYSTAL GROWTH 502 76 - 82 2018.11
BpeB, a major resistance-nodulation-cell division transporter from Burkholderia cenocepacia: construct design, crystallization and preliminary structural analysis. Reviewed
Horikawa T, Hung LW, Kim HB, Shaya D, Kim CY, Terwilliger TC, Yamashita E, Aoki M, Okada U, Murakami S
Acta crystallographica. Section F, Structural biology communications 74 ( Pt 11 ) 710 - 716 2018.11
Naritaka Kobayashi, Mihoko Maruyama, Yoichiro Mori, Suguru Fukukita, Hiroaki Adachi, Kazufumi Takano, Satoshi Murakami, Hiroyoshi Matsumura, Tsuyoshi Inoue, Masashi Yoshimura, Seiichiro Nakabayashi, Yusuke Mori, Hiroshi Y. Yoshikawa
The Journal of Physical Chemistry C 122 ( 38 ) 21983 - 21990 2018.9
Structure of the γ-ε complex of cyanobacterial F1-ATPase reveals a suppression mechanism of the γ subunit on ATP hydrolysis in phototrophs. Reviewed
Murakami S, Kondo K, Katayama S, Hara S, Sunamura EI, Yamashita E, Groth G, Hisabori T
The Biochemical journal 475 ( 18 ) 2925 - 2939 2018.9
Improvement of metastable crystal of acetaminophen via control of crystal growth rate Reviewed
Kosuke Nii, Mihoko Maruyama, Shino Okada, Hiroaki Adachi, Kazufumi Takano, Satoshi Murakami, Hiroshi Y. Yoshikawa, Hiroyoshi Matsumura, Tsuyoshi Inoue, Masayuki Imanishi, Katsuo Tsukamoto, Masashi Yoshimura, Yusuke Mori
Applied Physics Express 11 ( 3 ) 035501 - 035501 2018.3
Crystal structure of tripartite-type ABC transporter MacB from Acinetobacter baumannii Reviewed
Ui Okada, Eiki Yamashita, Arthur Neuberger, Mayu Morimoto, Hendrik W. van Veen, Satoshi Murakami
NATURE COMMUNICATIONS 8 ( 1 ) 2017.11
Structure of the MacAB-TolC ABC-type tripartite multidrug efflux pump Reviewed
Anthony W. P. Fitzpatrick, Salome Llabres, Arthur Neuberger, James N. Blaza, Xiao-Chen Bai, Ui Okada, Satoshi Murakami, Hendrik W. van Veen, Ulrich Zachariae, Sjors H. W. Scheres, Ben F. Luisi, Dijun Du
NATURE MICROBIOLOGY 2 ( 7 ) 17070 2017.7
Crystallization of acetaminophen form II by plastic-ball-assisted ultrasonic irradiation Reviewed
Yoichiro Mori, Mihoko Maruyama, Yoshinori Takahashi, Hiroshi Y. Yoshikawa, Shino Okada, Hiroaki Adachi, Shigeru Sugiyama, Kazufumi Takano, Satoshi Murakami, Hiroyoshi Matsumura, Tsuyoshi Inoue, Masashi Yoshimura, Yusuke Mori
APPLIED PHYSICS EXPRESS 10 ( 2 ) 025501 2017.2
Metastable crystal growth of acetaminophen using solution-mediated phase transformation Reviewed
Yoichiro Mori, Mihoko Maruyama, Yoshinori Takahashi, Hiroshi Y. Yoshikawa, Shino Okada, Hiroaki Adachi, Shigeru Sugiyama, Kazufumi Takano, Satoshi Murakami, Hiroyoshi Matsumura, Tsuyoshi Inoue, Masashi Yoshimura, Yusuke Mori
Applied Physics Express 10 ( 1 ) 015501 - 015501 2017.1
Promotion of protein crystal growth by actively switching crystal growth mode via femtosecond laser ablation Reviewed
Yusuke Tominaga, Mihoko Maruyama, Masashi Yoshimura, Haruhiko Koizumi, Masaru Tachibana, Shigeru Sugiyama, Hiroaki Adachi, Katsuo Tsukamoto, Hiroyoshi Matsumura, Kazufumi Takano, Satoshi Murakami, Tsuyoshi Inoue, Hiroshi Y. Yoshikawa, Yusuke Mori
NATURE PHOTONICS 10 ( 11 ) 723 - + 2016.11
A crystallization technique for obtaining large protein crystals with increased mechanical stability using agarose gel combined with a stirring technique Reviewed
Mihoko Maruyama, Yuki Hayashi, Hiroshi Y. Yoshikawa, Shino Okada, Haruhiko Koizumi, Masaru Tachibana, Shigeru Sugiyama, Hiroaki Adachi, Hiroyoshi Matsumura, Tsuyoshi Inoue, Kazufumi Takano, Satoshi Murakami, Masashi Yoshimura, Yusuke Mori
JOURNAL OF CRYSTAL GROWTH 452 172 - 178 2016.10
Molecular mechanism underlying promiscuous polyamine recognition by spermidine acetyltransferase. International journal
Shigeru Sugiyama, Sae Ishikawa, Hideyuki Tomitori, Mayumi Niiyama, Mika Hirose, Yuma Miyazaki, Kyohei Higashi, Michio Murata, Hiroaki Adachi, Kazufumi Takano, Satoshi Murakami, Tsuyoshi Inoue, Yusuke Mori, Keiko Kashiwagi, Kazuei Igarashi, Hiroyoshi Matsumura
The international journal of biochemistry & cell biology 76 87 - 97 2016.7
Protein crystallization with paper Reviewed
Miki Matsuoka, Keisuke Kakinouchi, Hiroaki Adachi, Mihoko Maruyama, Shigeru Sugiyama, Satoshi Sano, Hiroshi Y. Yoshikawa, Yoshinori Takahashi, Masashi Yoshimura, Hiroyoshi Matsumura, Satoshi Murakami, Tsuyoshi Inoue, Yusuke Mori, Kazufumi Takano
JAPANESE JOURNAL OF APPLIED PHYSICS 55 ( 5 ) 2016.5
Growth of high-strength protein crystals with nanofibers Reviewed
Miki Matsuoka, Keisuke Kakinouchi, Hiroaki Adachi, Mihoko Maruyama, Shigeru Sugiyama, Iori Nakabayashi, Hiroshi Tsuchikura, Atsushi Kuwahara, Satoshi Sano, Hiroshi Y. Yoshikawa, Yoshinori Takahashi, Masashi Yoshimura, Hiroyoshi Matsumura, Satoshi Murakami, Tsuyoshi Inoue, Yusuke Mori, Kazufumi Takano
APPLIED PHYSICS EXPRESS 9 ( 3 ) 2016.3
Structure, mechanism and cooperation of bacterial multidrug transporters Reviewed
Dijun Du, Hendrik W. van Veen, Satoshi Murakami, Klaas M. Pos, Ben F. Luisi
CURRENT OPINION IN STRUCTURAL BIOLOGY 33 76 - 91 2015.8
Selective crystallization of metastable phase of acetaminophen by ultrasonic irradiation Reviewed
Yoichiro Mori, Mihoko Maruyama, Yoshinori Takahashi, Kenji Ikeda, Suguru Fukukita, Hiroshi Y. Yoshikawa, Shino Okada, Hiroaki Adachi, Shigeru Sugiyama, Kazufumi Takano, Satoshi Murakami, Hiroyoshi Matsumura, Tsuyoshi Inoue, Masashi Yoshimura, Yusuke Mori
APPLIED PHYSICS EXPRESS 8 ( 6 ) 065501 2015.6
Spiral Growth Can Enhance Both the Normal Growth Rate and Quality of Tetragonal Lysozyme Crystals Grown under a Forced Solution Flow Reviewed
Yuki Hayashi, Mihoko Maruyama, Masashi Yoshimura, Shino Okada, Hiroshi Y. Yoshikawa, Shigeru Sugiyama, Hiroaki Adachi, Hiroyoshi Matsumura, Tsuyoshi Inoue, Kazufumi Takano, Satoshi Murakami, Yusuke Mori
CRYSTAL GROWTH & DESIGN 15 ( 5 ) 2137 - 2143 2015.5
Selective crystallization of the metastable phase of indomethacin at the interface of liquid/air bubble induced by femtosecond laser irradiation Reviewed
Kenji Ikeda, Mihoko Maruyama, Yoshinori Takahashi, Yoichiro Mori, Hiroshi Y. Yoshikawa, Shino Okada, Hiroaki Adachi, Shigeru Sugiyama, Kazufumi Takano, Satoshi Murakami, Hiroyoshi Matsumura, Tsuyoshi Inoue, Masashi Yoshimura, Yusuke Mori
APPLIED PHYSICS EXPRESS 8 ( 4 ) 045501 2015.4
Preliminary X-ray analysis of the binding domain of the soybean vacuolar sorting receptor complexed with a sorting determinant of a seed storage protein Reviewed
Nobuyuki Maruyama, Tomohiro Goshi, Shigeru Sugiyama, Mayumi Niiyama, Hiroaki Adachi, Kazufumi Takano, Satoshi Murakami, Tsuyoshi Inoue, Yusuke Mori, Hiroyoshi Matsumura, Bunzo Mikami
ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY COMMUNICATIONS 71 ( 2 ) 132 - 135 2015.2
Water-Mediated Recognition of Simple Alkyl Chains by Heart-Type Fatty-Acid-Binding Protein Reviewed
Shigeru Matsuoka, Shigeru Sugiyama, Daisuke Matsuoka, Mika Hirose, Sebastien Lethu, Hikaru Ano, Toshiaki Hara, Osamu Ichihara, S. Roy Kimura, Satoshi Murakami, Hanako Ishida, Eiichi Mizohata, Tsuyoshi Inoue, Michio Murata
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION 54 ( 5 ) 1508 - 1511 2015.1
Development of protein seed crystals reinforced with high-strength hydrogels Reviewed
Shigeru Sugiyama, Noriko Shimizu, Keisuke Kakinouchi, Osamu Hiraoka, Hiroyoshi Matsumura, Hiroshi Y. Yoshikawa, Yoshinori Takahashi, Mihoko Maruyama, Masashi Yoshimura, Hiroaki Adachi, Kazufumi Takano, Satoshi Murakami, Tsuyoshi Inoue, Michio Murata, Yusuke Mori
CRYSTENGCOMM 17 ( 42 ) 8064 - 8071 2015
A new practical technique for high quality protein crystallization with the solution stirring technique at the interface between high-concentrated hydrogel and solution Reviewed
Yusuke Aoki, Mihoko Maruyama, Yoshinori Takahashi, Masashi Yoshimura, Hiroshi Y. Yoshikawa, Shigeru Sugiyama, Hiroaki Adachi, Kazufumi Takano, Satoshi Murakami, Hiroyoshi Matsumura, Tsuyoshi Inoue, Yusuke Mori
JAPANESE JOURNAL OF APPLIED PHYSICS 53 ( 6 ) 065502 2014.6
β-Lactam Selectivity of Multidrug Transporters AcrB and AcrD Resides in the Proximal Binding Pocket Reviewed
Naoki Kobayashi, Norihisa Tamura, Hendrik W. van Veen, Akihito Yamaguchi, Satoshi Murakami
Journal of Biological Chemistry 289 ( 15 ) 10680 - 10690 2014.4
Crystallization and preliminary crystallographic studies of PotA, a membrane-associated ATPase of the spermidine-preferential uptake system in Thermotoga maritima Reviewed
Shigeru Sugiyama, Keiko Kashiwagi, Keisuke Kakinouchi, Hideyuki Tomitori, Ken Kanai, Michio Murata, Hiroaki Adachi, Hiroyoshi Matsumura, Kazufumi Takano, Satoshi Murakami, Tsuyoshi Inoue, Yusuke Mori, Kazuei Igarashi
Acta Crystallographica Section F:Structural Biology Communications 70 ( 6 ) 738 - 741 2014
Laser ablation for protein crystal nucleation and seeding Reviewed
Hiroshi Y. Yoshikawa, Ryota Murai, Hiroaki Adachi, Shigeru Sugiyama, Mihoko Maruyama, Yoshinori Takahashi, Kazufumi Takano, Hiroyoshi Matsumura, Tsuyoshi Inoue, Satoshi Murakami, Hiroshi Masuhara, Yusuke Mori
CHEMICAL SOCIETY REVIEWS 43 ( 7 ) 2147 - 2158 2014
Structure of the human-heart fatty-acid-binding protein 3 in complex with the fluorescent probe 1-anilinonaphthalene-8-sulphonic acid Reviewed
Mika Hirose, Shigeru Sugiyama, Hanako Ishida, Mayumi Niiyama, Daisuke Matsuoka, Toshiaki Hara, Eiichi Mizohata, Satoshi Murakami, Tsuyoshi Inoue, Shigeru Matsuoka, Michio Murata
JOURNAL OF SYNCHROTRON RADIATION 20 923 - 928 2013.11
Expression, purification, crystallization and preliminary crystallographic analysis of spermidine acetyltransferase from Escherichia coli. International journal
Mayumi Niiyama, Shigeru Sugiyama, Mika Hirose, Sae Ishikawa, Hideyuki Tomitori, Kyohei Higashi, Tomoko Yamashita, Hiroaki Adachi, Kazufumi Takano, Satoshi Murakami, Michio Murata, Tsuyoshi Inoue, Yusuke Mori, Keiko Kashiwagi, Hiroyoshi Matsumura, Kazuei Igarashi
Acta crystallographica. Section F, Structural biology and crystallization communications 69 ( Pt 8 ) 884 - 7 2013.8
Protein Crystal Growth under a Forced Solution Flow
丸山美帆子, 中村真利子, 佐崎元, 高橋義典, 吉村政志, 吉川洋史, 杉山成, 安達宏昭, 松村浩由, 井上豪, 高野和文, 村上聡, 森勇介
日本結晶成長学会誌 40 ( 2 ) 72 - 82 2013.7
A Novel Approach for Protein Crystallization by a Synthetic Hydrogel with Thermoreversible Gelation Polymer Reviewed
Shigeru Sugiyama, Noriko Shimizu, Gen Sazaki, Mika Hirose, Yoshinori Takahashi, Mihoko Maruyama, Hiroyoshi Matsumura, Hiroaki Adachi, Kazufumi Takano, Satoshi Murakami, Tsuyoshi Inoue, Yusuke Mori
CRYSTAL GROWTH & DESIGN 13 ( 5 ) 1899 - 1904 2013.5
Xin-Qiu Yao, Nobuhiro Kimura, Satoshi Murakami, Shoji Takada
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY 135 ( 20 ) 7474 - 7485 2013.5
Effect of Gel-Solution Interface on Femtosecond Laser-Induced Nucleation of Protein Reviewed
Satoshi Nakayama, Hiroshi Y. Yoshikawa, Ryota Murai, Masateru Kurata, Mihoko Maruyama, Shigeru Sugiyama, Yusuke Aoki, Yoshinori Takahashi, Masashi Yoshimura, Seiichiro Nakabayashi, Hiroaki Adachi, Hiroyoshi Matsumura, Tsuyoshi Inoue, Kazufumi Takano, Satoshi Murakami, Yusuke Mori
CRYSTAL GROWTH & DESIGN 13 ( 4 ) 1491 - 1496 2013.4
Growth of Protein Crystals in Hydrogels with High Strength
SUGIYAMA Shigeru, HIROSE Mika, SAZAKI Gen, ADACHI Hiroaki, TAKANO Kazufumi, MURAKAMI Satoshi, MATSUMURA Hiroyoshi, MARUYAMA Mihoko, INOUE Tsuyoshi, MORI Yusuke
Nihon Kessho Gakkaishi 54 ( 5 ) 300 - 303 2012.10
Spatially Precise, Soft Microseeding of Single Protein Crystals by Femtosecond Laser Ablation Reviewed
Hiroshi Y. Yoshikawa, Yoichiroh Hosokawa, Ryota Murai, Gen Sazaki, Tomoya Kitatani, Hiroaki Adachi, Tsuyoshi Inoue, Hiroyoshi Matsumura, Kazufumi Takano, Satoshi Murakami, Seiichiro Nakabayashi, Yusuke Mori, Hiroshi Masuhara
CRYSTAL GROWTH & DESIGN 12 ( 9 ) 4334 - 4339 2012.9
Effects of a Forced Solution Flow on the Step Advancement on {110} Faces of Tetragonal Lysozyme Crystals: Direct Visualization of Individual Steps under a Forced Solution Flow Reviewed
Mihoko Maruyama, Hisato Kawahara, Gen Sazaki, Syou Maki, Yoshinori Takahashi, Hiroshi Y. Yoshikawa, Shigeru Sugiyama, Hiroaki Adachi, Kazufumi Takano, Hiroyoshi Matsumura, Tsuyoshi Inoue, Satoshi Murakami, Yusuke Mori
CRYSTAL GROWTH & DESIGN 12 ( 6 ) 2856 - 2863 2012.6
Growth of Protein Crystals in Hydrogels Prevents Osmotic Shock Reviewed
Shigeru Sugiyama, Mihoko Maruyama, Gen Sazaki, Mika Hirose, Hiroaki Adachi, Kazufumi Takano, Satoshi Murakami, Tsuyoshi Inoue, Yusuke Mori, Hiroyoshi Matsumura
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY 134 ( 13 ) 5786 - 5789 2012.4
Crystallization and preliminary X-ray crystallographic analysis of a helicase-like domain from a tomato mosaic virus replication protein Reviewed
Hongyu Xiang, Mayumi Niiyama, Shigeru Sugiyama, Hiroaki Adachi, Kazufumi Takano, Satoshi Murakami, Tsuyoshi Inoue, Yusuke Mori, Masayuki Ishikawa, Hiroyoshi Matsumura, Etsuko Katoh
ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY COMMUNICATIONS 67 ( 12 ) 1649 - 1652 2011.12
Protein Nucleation Triggered by Femtosecond Laser-Induced Cavitation Bubbles
吉川洋史, 村井良多, 杉山成, 安達宏昭, 松村浩由, 井上豪, 村上聡, 高野和文, 森勇介
日本結晶成長学会誌 38 ( 3 ) 161 - 168 2011.10
Influence of energy and wavelength on femtosecond laser-induced nucleation of protein Reviewed
Ryota Murai, Hiroshi Y. Yoshikawa, Hitoshi Hasenaka, Yoshinori Takahashi, Mihoko Maruyama, Shigeru Sugiyama, Hiroaki Adachi, Kazufumi Takano, Hiroyoshi Matsumura, Satoshi Murakami, Tsuyoshi Inoue, Yusuke Mori
CHEMICAL PHYSICS LETTERS 510 ( 1-3 ) 139 - 142 2011.6
Laser-induced nucleation in protein crystallization: Local increase in protein concentration induced by femtosecond laser irradiation
Natsuko Iefuji, Ryota Murai, Mihoko Maruyama, Yoshinori Takahashi, Shigeru Sugiyama, Hiroaki Adachi, Hiroyoshi Matsumura, Satoshi Murakami, Tsuyoshi Inoue, Yusuke Mori, Yuichi Koga, Kazufumi Takano, Shigenori Kanaya
Journal of Crystal Growth 318 ( 1 ) 741 - 744 2011.3
Effect of Evaporation on Protein Crystals Grown in Semi-Solid Agarose Hydrogel Reviewed
Shigeru Sugiyama, Mika Hirose, Noriko Shimizu, Mayumi Niiyama, Mihoko Maruyama, Gen Sazaki, Ryota Murai, Hiroaki Adachi, Kazufumi Takano, Satoshi Murakami, Tsuyoshi Inoue, Yusuke Mori, Hiroyoshi Matsumura
JAPANESE JOURNAL OF APPLIED PHYSICS 50 ( 2 ) 025502 (4 pages) 2011.2
Approach for growth of high-quality and large protein crystals Reviewed
Hiroyoshi Matsumura, Shigeru Sugiyama, Mika Hirose, Keisuke Kakinouchi, Mihoko Maruyama, Ryota Murai, Hiroaki Adachi, Kazufumi Takano, Satoshi Murakami, Yusuke Mori, Tsuyoshi Inoue
JOURNAL OF SYNCHROTRON RADIATION 18 ( 1 ) 16 - 19 2011.1
Drug export and allosteric coupling in a multidrug transporter revealed by molecular simulations Reviewed
Xin-Qiu Yao, Hiroo Kenzaki, Satoshi Murakami, Shoji Takada
NATURE COMMUNICATIONS 1 117 2010.11
Conformational plasticity of RNA for target recognition as revealed by the 2.15 angstrom crystal structure of a human IgG-aptamer complex Reviewed
Yusuke Nomura, Shigeru Sugiyama, Taiichi Sakamoto, Shin Miyakawa, Hiroaki Adachi, Kazufumi Takano, Satoshi Murakami, Tsuyoshi Inoue, Yusuke Mori, Yoshikazu Nakamura, Hiroyoshi Matsumura
NUCLEIC ACIDS RESEARCH 38 ( 21 ) 7822 - 7829 2010.11
Estimated effects of silicone glue on protein crystal growth
Mihoko Maruyama, Noriko Shimizu, Shigeru Sugiyama, Yoshinori Takahashi, Hiroaki Adachi, Kazufumi Takano, Satoshi Murakami, Tsuyoshi Inoue, Hiroyoshi Matsumura, Yusuke Mori
JOURNAL OF CRYSTAL GROWTH 312 ( 19 ) 2771 - 2774 2010.9
Growth of large protein crystals by a large-scale hanging-drop method Reviewed
Keisuke Kakinouchi, Tsutomu Nakamura, Taro Tamada, Hiroaki Adachi, Shigeru Sugiyama, Mihoko Maruyama, Yoshinori Takahashi, Kazufumi Takano, Satoshi Murakami, Tsuyoshi Inoue, Ryota Kuroki, Yusuke Mori, Hiroyoshi Matsumura
JOURNAL OF APPLIED CRYSTALLOGRAPHY 43 ( 4 ) 937 - 939 2010.8
Crystal Growth Procedure of HIV-1 Protease-Inhibitor KNI-272 Complex for Neutron Structural Analysis at 1.9 angstrom Resolution Reviewed
Noriko Shimizu, Shigeru Sugiyama, Mihoko Maruyama, Yoshinori Takahashi, Motoyasu Adachi, Taro Tamada, Koushi Hidaka, Yoshio Hayashi, Tooru Kimura, Yoshiaki Kiso, Hiroaki Adachi, Kazufumi Takano, Satoshi Murakami, Tsuyoshi Inoue, Ryota Kuroki, Yusuke Mori, Hiroyoshi Matsumura
CRYSTAL GROWTH & DESIGN 10 ( 7 ) 2990 - 2994 2010.7
Molecular resolution investigation of tetragonal lysozyme (110) face in liquid by frequency-modulation atomic force microscopy Reviewed
Ken Nagashima, Masayuki Abe, Seizo Morita, Noriaki Oyabu, Kei Kobayashi, Hirofumi Yamada, Masahiro Ohta, Ryohei Kokawa, Ryota Murai, Hiroyoshi Matsumura, Hiroaki Adachi, Kazufumi Takano, Satoshi Murakami, Tsuyoshi Inoue, Yusuke Mori
JOURNAL OF VACUUM SCIENCE & TECHNOLOGY B 28 ( 3 ) 2010.5
Femtosecond laser processing of protein crystals grown in agarose gel Reviewed
Hitoshi Hasenaka, Shigeru Sugiyama, Mika Hirose, Noriko Shimizu, Tomoya Kitatani, Yoshinori Takahashi, Hiroaki Adachi, Kazufumi Takano, Satoshi Murakami, Tsuyoshi Inoue, Yusuke Mori, Hiroyoshi Matsumura
JOURNAL OF CRYSTAL GROWTH 312 ( 1 ) 73 - 78 2009.12
Promotion of Crystal Nucleation of Protein by Semi-Solid Agarose Gel Reviewed
Kana Tanabe, Mika Hirose, Ryota Murai, Shigeru Sugiyama, Noriko Shimizu, Mihoko Maruyama, Yoshinori Takahashi, Hiroaki Adachi, Kazufumi Takano, Satoshi Murakami, Yusuke Mori, Eiichi Mizohata, Tsuyoshi Inoue, Hiroyoshi Matsumura
APPLIED PHYSICS EXPRESS 2 ( 12 ) 125501 2009.12
Femtosecond Laser Processing of Agarose Gel Surrounding Protein Crystals for Development of an Automated Crystal Capturing System Reviewed
Shigeru Sugiyama, Hitoshi Hasenaka, Mika Hirose, Noriko Shimizu, Tomoya Kitatani, Yoshinori Takahashi, Hiroaki Adachi, Kazufumi Takano, Satoshi Murakami, Tsuyoshi Inoue, Yusuke Mori, Hiroyoshi Matsumura
JAPANESE JOURNAL OF APPLIED PHYSICS 48 ( 10 ) 105502 (5 pages) 2009.10
Protein Crystallization in Agarose Gel with High Strength: Developing an Automated System for Protein Crystallographic Processes Reviewed
Shigeru Sugiyama, Kana Tanabe, Mika Hirose, Tomoya Kitatani, Hitoshi Hasenaka, Yoshinori Takahashi, Hiroaki Adachi, Kazufumi Takano, Satoshi Murakami, Yusuke Mori, Tsuyoshi Inoue, Hiroyoshi Matsumura
JAPANESE JOURNAL OF APPLIED PHYSICS 48 ( 7 ) 075502 (5 pages) 2009.7
Dynamic nature of disulphide bond formation catalysts revealed by crystal structures of DsbB Reviewed
Kenji Inaba, Satoshi Murakami, Atsushi Nakagawa, Hiroka Iida, Mai Kinjo, Koreaki Ito, Mamoru Suzuki
EMBO JOURNAL 28 ( 6 ) 779 - 791 2009.3
Crystallization and preliminary X-ray crystallographic analysis of Ca(2+)-free primary Ca(2+)-sensor of Na(+)/Ca(2+) exchanger Reviewed
Masashi Mima, Chika Kawai, Kumsun Paku, Koji Tomoo, Toshimasa Ishida, Shigeru Sugiyama, Hiroyoshi Matsumura, Tomoya Kitatani, Hiroshi Y. Yoshikawa, Syou Maki, Hiroaki Adachi, Kazufumi Takano, Satoshi Murakami, Tsuyoshi Inoue, Yusuke Mori, Satomi Kita, Takahiro Iwamoto
ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY AND CRYSTALLIZATION COMMUNICATIONS 64 ( 12 ) 1125 - 1127 2008.12
Crystallization and preliminary X-ray diffraction studies of an RNA aptamer in complex with the human IgG Fc fragment Reviewed
Shigeru Sugiyama, Yusuke Nomura, Taiichi Sakamoto, Tomoya Kitatani, Asako Kobayashi, Shin Miyakawa, Yoshinori Takahashi, Hiroaki Adachi, Kazufumi Takano, Satoshi Murakami, Tsuyoshi Inoue, Yusuke Mori, Yoshikazu Nakamura, Hiroyoshi Matsumura
ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY AND CRYSTALLIZATION COMMUNICATIONS 64 ( 10 ) 942 - 944 2008.10
Tsuyoshi Inoue, Hiroaki Adachi, Satoshi Murakami, Kazufumi Takano, Hiroyoshi Matsumura, Yusuke Mori, Yoshifumi Fukunishi, Haruki Nakamura, Takayoshi Kinoshita, Isao Nakanishi, Yasushi Okuno, Satoshi Minakata, Shinji Shimojo, Tsuneaki Sakata
Yakugaku Zasshi 128 ( 4 ) 497 - 505 2008.4
New technique of manipulating a protein crystal using adhesive material Reviewed
Tomoya Kitatani, Shigeru Sugiyama, Hiroyoshi Matsumura, Hiroaki Adachi, Hiroshi Y. Yoshikawa, Syou Maki, Satoshi Murakami, Tsuyoshi Inoue, Yusuke Mori, Kazufumi Takano
APPLIED PHYSICS EXPRESS 1 ( 3 ) 037002 (3 pages) 2008.3
2P-119 X-ray structure of RNA aptamer in complex with human immunoglobulin G(The 46th Annual Meeting of the Biophysical Society of Japan)
Sugiyama Shigeru, Matsumura Hiroyoshi, Kitatani Tomoya, Nomura Yusuke, Sakamoto Taiichi, Miyakawa Shin, Nakamura Yoshikazu, Takahashi Yoshinori, Adachi Hiroaki, Takano Kazufumi, Murakami Satoshi, Inoue Tsuyoshi, Mori Yusuke
Seibutsu Butsuri 48 S93 2008
Structural basis of xenobiotic recognition by a bacterial xenobiotic exporter Reviewed
Satoshi Murakami, Akihito Yamaguchi
SEIKAGAKU 79 ( 6 ) 542 - 549 2007.6
Crystal structure of the DsbB-DsbA complex reveals a mechanism of disulfide bond generation Reviewed
Kenji Inaba, Satoshi Murakami, Mamoru Suzuki, Atsushi Nakagawa, Eiki Yamashita, Kengo Okada, Koreaki Ito
CELL 127 ( 4 ) 789 - 801 2006.11
Crystal structures of a multidrug transporter reveal a functionally rotating mechanism Reviewed
Satoshi Murakami, Ryosuke Nakashima, Eiki Yamashita, Takashi Matsumoto, Akihito Yamaguchi
NATURE 443 ( 7108 ) 173 - 179 2006.9
Tomoya Tsukazaki, Hiroyuki Mori, Shuya Fukai, Tomoyuki Numata, Anna Perederina, Hiroaki Adachi, Hiroyoshi Matsumura, Kazufumi Takano, Satoshi Murakami, Tsuyoshi Inoue, Yusuke Mori, Takatomo Sasaki, Dmitry G. Vassylyev, Osamu Nureki, Koreaki Ito
Acta Crystallographica Section F Structural Biology and Crystallization Communications 62 ( 4 ) 376 - 380 2006.4
Temperature-Screening System for Determining Protein Crystallization Conditions
Hiroaki Adachi, Ai Niino, Kazufumi Takano, Hiroyoshi Matsumura, Satoshi Murakami, Tsuyoshi Inoue, Yusuke Mori, Takatomo Sasaki
Japanese Journal of Applied Physics 44 ( 6A ) 4080 - 4083 2005.6
Protein Cryocrystallography Using Laser-Processed Crystal
Hiroshi Kitano, Hiroyoshi Matsumura, Hiroaki Adachi, Satoshi Murakami, Kazufumi Takano, Tsuyoshi Inoue, Yusuke Mori, Masaaki Doi, Takatomo Sasaki
Japanese Journal of Applied Physics 44 ( 2 ) L54 - L56 2005.1
3P165 Improvement of ATPsynthase crystals
Shirakihara Y., Shiratori A., Murakami S., Adachi H., Matumura H., Takano K., Inoue G., Moti Y., Sasaki T., Suzuki T., Yoshida M.
Seibutsu Butsuri 45 S245 2005
Functional analysis of bacterial efflux transporter AcrB based on its crystal structure Reviewed
S Murakami, A Yamaguchi
YAKUGAKU ZASSHI-JOURNAL OF THE PHARMACEUTICAL SOCIETY OF JAPAN 125 66 - 67 2005
Membrane Protein Crystallization Using Laser Irradiation
Hiroaki Adachi, Satoshi Murakami, Ai Niino, Hiroyoshi Matsumura, Kazufumi Takano, Tsuyoshi Inoue, Yusuke Mori, Akihito Yamaguchi, Takatomo Sasaki
Japanese Journal of Applied Physics 43 ( No. 10B ) L1376 - L1378 2004.10
Structure and function of bacterial multi-drug efflux transporter AcrB Reviewed
S Murakami, A Yamaguchi
YAKUGAKU ZASSHI-JOURNAL OF THE PHARMACEUTICAL SOCIETY OF JAPAN 124 177 - 180 2004
Identification of the substrate-recognition site in the xenobiotic exporter AcrB using AcrB/AcrD chimeric proteins Reviewed
N Kobayashi, N Tamura, S Murakami, A Yamaguchi
YAKUGAKU ZASSHI-JOURNAL OF THE PHARMACEUTICAL SOCIETY OF JAPAN 124 ( Suppl. 4 ) 325 - 328 2004
Multidrug-exporting secondary transporters Reviewed
S Murakami, A Yamaguchi
CURRENT OPINION IN STRUCTURAL BIOLOGY 13 ( 4 ) 443 - 452 2003.8
Crystal structure of bacterial multidrug efflux transporter AcrB Reviewed
S Murakami, R Nakashima, E Yamashita, A Yamaguchi
NATURE 419 ( 6907 ) 587 - 593 2002.10
凝固ゲルに代わる高強度タンパク質結晶育成法
松岡美紀, 垣之内啓介, 垣之内啓介, 安達宏昭, 安達宏昭, 丸山美帆子, 杉山成, 杉山成, 中林伊織, 土倉弘至, 桑原厚司, 佐野智, 吉川洋史, 高橋義典, 吉村政志, 松村浩由, 松村浩由, 村上聡, 村上聡, 井上豪, 井上豪, 森勇介, 森勇介, 高野和文, 高野和文
日本結晶学会年会講演要旨集 2015 2015
不織布を用いたタンパク質結晶育成法
松岡美紀, 垣之内啓介, 垣之内啓介, 安達宏昭, 安達宏昭, 丸山美帆子, 杉山成, 杉山成, 佐野智, 吉川洋史, 高橋義典, 吉村政志, 松村浩由, 松村浩由, 村上聡, 村上聡, 井上豪, 井上豪, 森勇介, 森勇介, 高野和文, 高野和文
結晶成長国内会議予稿集(CD-ROM) 45th 2015
緑膿菌由来多剤排出トランスポーターMexBの構造生物学的研究
米原涼, 山下栄樹, 原田健一, 佐藤尚紀, 松浦孝範, 村上聡, 中江太治, 中川敦史
日本蛋白質科学会年会プログラム・要旨集 11th 120 2011.5
Growth of Protein Crystals by Syringe-Type Top-Seeded Solution Growth
Hiroyoshi Matsumura, Keisuke Kakiniuchi, Tsutomu Nakamura, Shigeru Sugiyama, Mihoko Maruyama, Hiroaki Adachi, Kazufumi Takano, Satoshi Murakami, Tsuyoshi Inoue, Yusuke Mori
CRYSTAL GROWTH & DESIGN 11 ( 5 ) 1486 - 1492 2011.5
緑膿菌におけるRND型トランスポーターMexBの高分解能X線結晶構造解析
米原涼, 山下栄樹, 原田健一, 佐藤尚紀, 松浦孝範, 鈴木守, 村上聡, 中江太治, 中川敦史
日本放射光学会年会・放射光科学合同シンポジウム予稿集 24th 112 2011.1
Crystal structure analysis of multidrug transporter MexB
Ryo Yonehara, Eiki Yamashita, Kenichi Harada, Naoki Sato, Takanori Matsuura, Satoshi Murakami, Taiji Nakae, Atsushi Nakagawa
ACTA CRYSTALLOGRAPHICA A-FOUNDATION AND ADVANCES 67 C744 - C744 2011
緑膿菌の多剤排出ポンプ,MexA‐OprM複合体のX線結晶構造解析への試み
米原涼, 山下栄樹, 原田健一, 佐藤尚紀, 松浦孝範, 鈴木守, 村上聡, 中江太治, 中川敦史
日本蛋白質科学会年会プログラム・要旨集 10th 147 2010.5
INOUE Tsuyoshi, MATSUMURA Hiroyoshi, ADACHI Hiroaki, MORI Yusuke, TAKANO Kazufumi, MURAKAMI Satoshi, FUKUNISHI Yoshifumi, NAKAMURA Haruki, KINOSHITA Takayoshi, NAKANISHI Isao, OKUNO Yasushi, MINAKATA Satoshi, MIKAMI Yoshiaki, SAKUMA Toshihiro, KITAJIMA Masato, FUKUOKA Yoshitada, TAKADA Toshikazu, SAKATA Tsuneaki
Nihon Kessho Gakkaishi 52 ( 1 ) 89 - 94 2010.2
Enhancement of femtosecond laser-induced nucleation of protein in a gel solution
Ryota Murai, Hiroshi Y. Yoshikawa, Yoshinori Takahashi, Mihoko Maruyama, Shigeru Sugiyama, Gen Sazaki, Hiroaki Adachi, Kazufumi Takano, Hiroyoshi Matsumura, Satoshi Murakami, Tsuyoshi Inoue, Yusuke Mori
APPLIED PHYSICS LETTERS 96 ( 4 ) 2010.1
緑膿菌由来多剤排出トランスポーター,MexBの高分解能X線結晶構造解析
米原涼, 山下栄樹, 原田健一, 佐藤尚紀, 松浦孝範, 鈴木守, 村上聡, 中江太治, 中川敦史
日本結晶学会年会講演要旨集 2010 95 2010
2P007 Crystal structure of the Escherichia coli spermidine acetyl-transferase in complex with spermidine and coenzyme A(The 48th Annual Meeting of the Biophysical Society of Japan)
Ishikawa Sae, Sugiyama Shigeru, Miiyama Mayumi, Tomitori Hideyuki, Higashi Kyohei, Adachi Hiroaki, Takano Kazufumi, Murakami Satoshi, Mori Yusuke, Inoue Tsuyoshi, Kashiwagi Keiko, Igarashi Kazuei, Matsumura Hiroyoshi
Seibutsu Butsuri 50 ( 2 ) S83 2010
Growth of Large Protein Crystals by Top-Seeded Solution Growth Together with the Floating and Solution-Stirring Technique
Noriko Shimizu, Shigeru Sugiyama, Mihoko Maruyama, Hiroshi Y. Yoshikawa, Yoshinori Takahashi, Hiroaki Adachi, Kazufumi Takano, Satoshi Murakami, Tsuyoshi Inoue, Hiroyoshi Matsumura, Yusuke Mori
CRYSTAL GROWTH & DESIGN 9 ( 12 ) 5227 - 5232 2009.12
A Manipulating Tool for Protein Microcrystals in Solution Using Adhesive Materials
Tomoya Kitatani, Hiroaki Adachi, Shigeru Sugiyama, Hiroyoshi Matsumura, Ryota Murai, Yoshinori Takahashi, Satoshi Murakami, Tsuyoshi Inoue, Yusuke Mori, Kazufumi Takano
JAPANESE JOURNAL OF APPLIED PHYSICS 48 ( 11 ) 2009.11
Femtosecond laser-induced nucleation of protein in agarose gel
Hiroshi Y. Yoshikawa, Ryota Murai, Shigeru Sugiyama, Gen Sazaki, Tornoya Kitatani, Yoshinori Takahashi, Hiroaki Adachi, Hiroyoshi Matsumura, Satoshi Murakami, Tsuyoshi Inoue, Kazufumi Takano, Yusuke Mori
JOURNAL OF CRYSTAL GROWTH 311 ( 3 ) 956 - 959 2009.1
Laser energy dependence on femtosecond laser-induced nucleation of protein
Hiroshi Y. Yoshikawa, Ryota Murai, Syou Maki, Tomoya Kitatani, Shigeru Sugiyama, Gen Sazaki, Hiroaki Adachi, Tsuyoshi Inoue, Hiroyoshi Matsumura, Kazufumi Takano, Satoshi Murakami, Takatomo Sasaki, Yusuke Mori
APPLIED PHYSICS A-MATERIALS SCIENCE & PROCESSING 93 ( 4 ) 911 - 915 2008.12
Evaluation and Improvement of a Technique to Manipulate Protein Crystals in Solution
Tomoya Kitatani, Hiroaki Adachi, Shigeru Sugiyama, Hiroyoshi Matsumura, Ryota Murai, Yoshinori Takahashi, Satoshi Murakami, Tsuyshi Inoue, Yusuke Mori, Kazufumi Takano
JAPANESE JOURNAL OF APPLIED PHYSICS 47 ( 12 ) 8995 - 8997 2008.12
Crystallization and preliminary neutron diffraction studies of HIV-1 protease cocrystallized with inhibitor KNI-272
Hiroyoshi Matsumura, Motoyasu Adachi, Shigeru Sugiyama, Shino Okada, Megumi Yamakami, Taro Tamada, Koushi Hidaka, Yoshio Hayashi, Tooru Kimura, Yoshiaki Kiso, Tomoya Kitatani, Sho Maki, Hiroshi Y. Yoshikawa, Hiroaki Adachi, Kazufumi Takano, Satoshi Murakami, Tsuyoshi Inoue, Ryota Kuroki, Yusuke Mori
ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY AND CRYSTALLIZATION COMMUNICATIONS 64 1003 - 1006 2008.11
Protein crystallization in a 100 nl solution with new stirring equipment
S. Maki, R. Murai, H. Y. Yoshikawa, T. Kitatani, S. Nakata, H. Kawahara, H. Hasenaka, A. Kobayashi, S. Okada, S. Sugiyama, H. Adachi, H. Matsumura, K. Takano, S. Murakami, T. Inoue, T. Sasaki, Y. Mori
JOURNAL OF SYNCHROTRON RADIATION 15 269 - 272 2008.5
Effect of solution flow produced by rotary shaker on protein crystallization
Ryota Murai, Hiroshi Y. Yoshikawa, Hisato Kawahara, Syou Maki, Shigeru Sugiyama, Tomoya Kitatani, Hiroaki Adachi, Kazufumi Takano, Hiroyoshi Matsumura, Satoshi Murakami, Tsuyoshi Inoue, Takatomo Sasaki, Yusuke Mori
JOURNAL OF CRYSTAL GROWTH 310 ( 7-9 ) 2168 - 2172 2008.4
Growth of high-quality and large crystals of HIV protease for neutron crystallography
Megumi Yamakami, Hiroyoshi Matsumura, Shigeru Sugiyama, Shino Okada, Motoyasu Adachi, Taro Tamada, Ryota Kuroki, Koushi Hidaka, Yoshio Hayashi, Yoshiaki Kiso, Sho Maki, Tomoya Kitatani, Hiroshi Yoshikawa, Hiroaki Adachi, Kazufumi Takano, Satoshi Murakami, Tsuyoshi Inoue, Yusuke Mori
ACTA CRYSTALLOGRAPHICA A-FOUNDATION AND ADVANCES 64 C244 - C244 2008
Femtosecond laser-induced cleaving of protein crystal in water solution
Masafumi Kashii, Yoichiroh Hosokawa, Hiroshi Kitano, Hiroaki Adachi, Yusuke Mori, Kazufumi Takano, Hiroyoshi Matsumura, Tsuyoshi Inoue, Satoshi Murakami, Kazuomi Sugamoto, Hideki Yoshikawa, Takatomo Sasaki, Hiroshi Masuhara
APPLIED SURFACE SCIENCE 253 ( 15 ) 6447 - 6450 2007.5
Development of protein crystallization and processing: Femtosecond laser, all solid-state 193 nm laser, and solution stirring techniques
Yusuke Mori, Kazufurni Takano, Hiroaki Adachi, Tsuyoshi Inoue, Satoshi Murakami, Hiroyoshi Matsumura, Masafumi. Kashii, Hiroshi Y. Yoshikawa, Syou Maki, Tornoya Kitatani, Shino Okada, Takatomo Sasaki
COMMERCIAL AND BIOMEDICAL APPLICATIONS OF ULTRAFAST LASERS VII 6460 2007
Drug development value chain constructed by collaboration between the SOSHO project and the NPO BIOGRID
Tsuyoshi Inoue, Yuji Kado, Keiji Tokuoka, Hiroyoshi Matsumura, Yasushi Kai, Yusuke Mori, Hiroaki Adachi, Kazufumi Takano, Satoshi Murakami, Yoshifumi Fukunishi, Haruki Nakamura, Takayoshi Kinoshita, Isao Nakanishi, Yasushi Okuno, Seiji Minakata, Tsuneaki Sakata
AIP Conference Proceedings 902 85 - 88 2007
Application of femtosecond laser ablation for detaching grown protein crystals from glass capillary tube
Masafumi Kashii, Ryo Fujisawa, Hiroaki Adachi, Yusuke Mori, Takatomo Sasaki, Yuichi Koga, Kazufumi Takano, Shigenori Kanaya, Hiroyoshi Matsumura, Tsuyoshi Inoue, Satoshi Murakami, Kazuomi Sugamoto, Hideki Yoshikawa
JOURNAL OF BIOSCIENCE AND BIOENGINEERING 102 ( 4 ) 372 - 374 2006.10
Cooling-rate screening system for determining protein crystal growth conditions
Ryota Murai, Shinya Nakata, Masafumi Kashii, Hiroaki Adachi, Ai Niino, Kazufumi Takano, Hiroyoshi Matsumura, Satoshi Murakami, Tsuyoshi Inoue, Yusuke Mori, Takatomo Sasaki
JOURNAL OF CRYSTAL GROWTH 292 ( 2 ) 433 - 436 2006.7
Effect of ultrasonic irradiation on protein crystallization
Keisuke Kakinouchi, Hiroaki Adachi, Hiroyoshi Matsumura, Tsuyoshi Inoue, Satoshi Murakami, Yusuke Mori, Yuichi Koga, Kazufumi Takano, Shigenori Kanaya
JOURNAL OF CRYSTAL GROWTH 292 ( 2 ) 437 - 440 2006.7
Crystallization and preliminary crystallographic analysis of orotidine 5 '-monophosphate decarboxylase from the human malaria parasite Plasmodium falciparum
Sudaratana R. Krungkrai, Keiji Tokuoka, Yukiko Kusakari, Tsuyoshi Inoue, Hiroaki Adachi, Hiroyoshi Matsumura, Kazufumi Takano, Satoshi Murakami, Yusuke Mori, Yasushi Kai, Jerapan Krungkrai, Toshihiro Horii
ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY COMMUNICATIONS 62 542 - 545 2006.6
Crystallization and preliminary X-ray analysis of the tRNA thiolation enzyme MnmA from Escherichia coli complexed with tRNAGlu
Tomoyuki Numata, Yoshiho Ikeuchi, Shuya Fukai, Hiroaki Adachi, Hiroyoshi Matsumura, Kazufumi Takano, Satoshi Murakami, Tsuyoshi Inoue, Yusuke Mori, Takatomo Sasaki, Tsutomu Suzuki, Osamu Nureki
Acta Crystallographica Section F: Structural Biology and Crystallization Communications 62 ( 4 ) 368 - 371 2006.4
Solution-stirring method improves crystal quality of human triosephosphate isomerase
H Adachi, A Niino, T Kinoshita, M Warizaya, R Maruki, K Takano, H Matsumura, T Inoue, S Murakami, Y Mori, T Sasaki
JOURNAL OF BIOSCIENCE AND BIOENGINEERING 101 ( 1 ) 83 - 86 2006.1
Effect of laser irradiation on enzyme activity
S Murakami, M Kashii, H Kitano, H Adachi, K Takano, H Matsumura, T Inoue, Y Mori, M Doi, K Sugamoto, H Yoshikawa, T Sasaki
JAPANESE JOURNAL OF APPLIED PHYSICS PART 1-REGULAR PAPERS BRIEF COMMUNICATIONS & REVIEW PAPERS 44 ( 11 ) 8216 - 8218 2005.11
Semiautomatic protein crystallization system featuring crystallization solution preparation function
H Adachi, H Matsumura, T Inoue, A Niino, K Takano, S Murakami, Y Mori, T Sasaki
JAPANESE JOURNAL OF APPLIED PHYSICS PART 1-REGULAR PAPERS BRIEF COMMUNICATIONS & REVIEW PAPERS 44 ( 8 ) 6302 - 6303 2005.8
Processing of membrane protein crystal using ultraviolet laser irradiation
H Kitano, S Murakami, H Adachi, H Matsumura, K Takano, T Inoue, Y Mori, M Doi, T Sasaki
JOURNAL OF BIOSCIENCE AND BIOENGINEERING 100 ( 1 ) 50 - 53 2005.7
Protein crystal growth using laser-processed seed crystals
K Takeuchi, H Kitano, H Adachi, Y Mori, T Sasaki, H Matsumura, T Inoue, S Murakami, M Doi, Y Koga, K Takano, S Kanaya
JAPANESE JOURNAL OF APPLIED PHYSICS PART 1-REGULAR PAPERS BRIEF COMMUNICATIONS & REVIEW PAPERS 44 ( 5A ) 3177 - 3179 2005.5
Protein crystallization by combining laser irradiation and solution-stirring techniques
H Adachi, A Niino, S Murakami, K Takano, H Matsumura, T Kinoshita, M Warizaya, T Inoue, Y Mori, T Sasaki
JAPANESE JOURNAL OF APPLIED PHYSICS PART 1-REGULAR PAPERS BRIEF COMMUNICATIONS & REVIEW PAPERS 44 ( 3 ) 1365 - 1366 2005.3
Growth of protein crystal by temperature control
Ryota Murai, Shinya Nakata, Masafumi Kashii, Hiroaki Adachi, kazufumi Takano, Murakami Satoshi, Hiroyoshi Matsumura, Tsuyoshi Inoue, Yusuke Mori, Takatomo Sasaki
Journal of the Japanese Association for Crystal Growth 32 ( 3 ) 234 - 234 2005
Femtosecond laser processing of protein crystals in crystallization drop
M Kashii, H Kitano, Y Hosokawa, H Adachi, Y Mori, T Sasaki, H Masuhara, K Takano, H Matsumura, T Inoue, S Murakami, K Sugamoto, H Yoshikawa
JAPANESE JOURNAL OF APPLIED PHYSICS PART 2-LETTERS & EXPRESS LETTERS 44 ( 24-27 ) L873 - L875 2005
1SD02 Inhibitor design based on the crystal structure of prostaglandin synthase
Inoue T.
Seibutsu Butsuri 44 S7 2004
村上聡, 中島良介, 山下栄樹, 山口明人
日本分子生物学会年会プログラム・講演要旨集 26th 420 2003.11
Crystal Structure of Bacterial Multi-Drug Efflux Transporter AcrB
村上聡, 中島良介, 山下栄樹, 山口明人
日本結晶学会誌 45 ( 4 ) 256 - 261 2003.8
Crystal Structure of Bacterial Multi-drug Efflux Transporter AcrB
村上聡, 中島良介, 山下栄樹, 山口明人
放射光 16 ( 4 ) 204 - 212 2003.7
Crystal structure of bacterial multidrug efflux transporter AcrB
A Yamaguchi, S Murakami, R Nakshima, E Yamashita
FASEB JOURNAL 17 ( 5 ) A1185 - A1185 2003.3
村上聡, 中島良介, 山下栄樹, 山口明人
生体膜と薬物の相互作用シンポジウム講演要旨集 24th 63 - 66 2002.11
村上聡, 中島良介, 山下栄樹, 山口明人
日本生物物理学会年会講演予稿集 40th ( 2 ) S171 2002.11
村上聡, 中島良介, 山下栄樹, 山口明人
生化学 74 ( 8 ) 655 2002.8
文部科学大臣表彰 若手科学者賞
2007 文部科学省 多剤排出蛋白質の結晶構造に基づく作動メカニズムの研究
村上 聡
日経BP技術賞・大賞
2006 日経BP社
村上 聡
Structural and Functional analysis on tripartite ABC transporters.
Grant number：21H02412 2021.4 - 2024.3
Japan Society for the Promotion of Science Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)
Grant amount：\17420000 （ Direct Cost: \13400000 、 Indirect Cost：\4020000 ）
Molecular mechanisms underlying the selective membrane localization of bacterial lipoproteins
Grant number：18K05396 2018.4 - 2023.3
Japan Society for the Promotion of Science Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)
Grant amount：\4420000 （ Direct Cost: \3400000 、 Indirect Cost：\1020000 ）
Structural and Functional analysis on tripartite ABC transporters.
Grant number：18H02386 2018.4 - 2021.3
Japan Society for the Promotion of Science Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)
MURAKAMI SATOSHI
Grant amount：\17290000 （ Direct Cost: \13300000 、 Indirect Cost：\3990000 ）
We solved a high-resolution crystal structure of MacB homolog. Compared to the published MacB structure, we observed several structural differences between them, especially around the transmembrane and the periplasmic domain. Comparison of these structures provides new insight on how substrates in the periplasm are captured and transported through the connecting pathway. In addition, we found that the substrate transport mechanism from the periplasmic space is similar to that of RND transporters, which we are studying separately. In view of the similarities and differences in substrate transport mechanisms, we proposed the “periplasmic alternating-access mechanism”. We also helped to propose a new classification of ABC transporters by the ABC transporter research society. And MacB is newly categolised as the type-VII ABC transporter.
Opening up New Structural Biology by High-speed AFM
Grant number：24227005 2012.5 - 2017.3
Japan Society for the Promotion of Science Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (S)
Ando Toshio, UCHIHASHI Takayuki, FUKUMORI Yoshihiro, FUKUMA Takeshi, KODERA Noriyuki, KONNO Hiroki, WONG Richard, MURAKAMI Satoshi, OGURA Teru, TOYOSHIMA Yoko, KANDORI Hideki
Grant amount：\215540000 （ Direct Cost: \165800000 、 Indirect Cost：\49740000 ）
We worked on three subjects. In subject 1, we elucidated the functional mechanism of various protein systems by observing their dynamic processes using high-speed AFM (HS-AFM) that we had established before. Besides, we demonstrated that HS-AFM can analyze the structure of intrinsically disordered proteins that are difficult to analyze with conventional methods. In subject 2, we developed fast/wide-area scanning techniques without generation of unwanted vibrations and an interactive HS-AFM technique that can manipulate the sample during imaging. The usefulness of these new techniques were demonstrated. In addition, we developed cantilever-scan type of HS-AFM combined with fluorescence microscopy, and thereby, made it possible to capture HS-AFM and fluorescence images simultaneously. In subject 3, we developed high-speed scanning ion conductance microscopy that can image the sample without contact with the sample. The imaging speed reached a level 100-times faster than before.
Structural and Functional analysis of RND type transporter
Grant number：19207009 2007 - 2009
Japan Society for the Promotion of Science Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (A)
MURAKAMI Satoshi
Grant amount：\42380000 （ Direct Cost: \32600000 、 Indirect Cost：\9780000 ）
Structure of AcrB was improved beyond 2.5Å resolution.
Along the substrate translocation pathway of AcrB, an important region where determine the substrate specificity was found.
Drug transport across the membrane was observed for Gram-positive RND transporter.
Structure, rotation and regulation of ATP synthase(FoF1)
Grant number：18107004 2006 - 2010
Japan Society for the Promotion of Science Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (S)
YOSHIDA Masasuke, HISABORI Toru, MURAKAMI Satoshi
Grant amount：\111280000 （ Direct Cost: \85600000 、 Indirect Cost：\25680000 ）
We made F_1-ATPases containing one or two mutant. β subunits that have the altered catalytic kinetics. The analysis of their asymmetric stepwise rotations reveal that, when watching one. β subunit in F_1-ATPase, it binds ATP at 0°, cleaves ATP after～ 200°rotation, and undergoes the final catalytic event, presumably the product release, after～ 320°rotation. Three. β subunits carry out this cycle at 120°difference like "trolling a tune" and their cyclic inter-domain bending motions would induce the rotation of γ subunit.
Rotation of the central shaft γ subunit in a molecular motor F_1-ATPase is assumed to correlate with and probably be driven by domain motions of the three catalytic. β subunits. Here we observe directly these. β motions through an attached fluorophore, concomitantly with 80 degrees and 40 degrees substep rotations of γ in the same single molecules. We show the sequence of conformations that each β subunit undergoes in three-step bending, an approximately 40 degrees counterclockwise turn followed by two approximately 20 degrees clockwise turns, occurring in synchronization with two substep rotations of γ. The results indicate that most previous crystal structures mimic the conformational set of three β subunits in the catalytic dwells. Moreover, a previously undescribed set of β conformations, open, closed and partially closed, is revealed in the ATP-waiting dwells. The present study thus bridges the gap between the chemical and mechanical steps in F_1-ATPase.
多剤排出トランスポーター群の内因性基質の解析
Grant number：17770091 2005 - 2007
日本学術振興会 科学研究費助成事業 若手研究(B)
村上 聡
Grant amount：\2700000 （ Direct Cost: \2700000 ）
多剤排出トランスポーター群は構造的に関連性の乏しい多様な生体にとって異物となる化合物を排出する膜蛋白質群である。これは、古細菌からヒトの細菌に至るまで広く生物界に存在し、あらゆる細胞の持つ最も基本的な生体防御機構であると理解されている。AcrBは大腸菌で構成的に発現しているトランスポーターで、広い基質選択性を持ち、大腸菌の最も強力な排出トランスポーターであり、また薬剤自然抵抗性の主因としても知られている。我々はこの立体構造を2002年に解き、その後基質認識機構や、薬剤排出機構の解明を進める一方で、本助成のテーマであるこれらトランスポーターの発現を制御する転写制御因子の結晶化および構造解析を進めてきた。この転写因子は、細胞内に進入した薬剤を検地するセンサでもあり、多剤認識メカニズムについては、その制御対象であるトランスポーターと比較することは非常に興味深い。本年の研究成果として以下の項目を挙げる。
1)AcrRの1.1Å分解能での超高分解能X腺結晶構造解析に成功した。(論文準備中)
2)結晶標品からクロロホルム・メタノール法で脂質成分を抽出し、ガスクロマトグラフィーを用いたマス・スペクトログラフィー(GCマス)による分析により、抽出した脂溶性リガンドの定性を行った。その結果、いわゆる脂溶性物質の混合物として検出した。
GCマスの解析結果として現れた脂溶性リガンドのうち、結晶構造中に見出された内因性基質の電子密度の形と部分的に合致するものもあり、今後より詳細なマス解析を行うと共に、バイオ・アッセイを行い、生物学的意味づけを行うことで生理的基質の同定を行う。
また大腸菌を使って、内因性基質がいかなる原因で発生するのか?発生したその物質は細胞にとってどれほど有害なものか?それはどのような遺伝子群を高進させるか?いかなる種類のトランスポーターがその排除にあたるか。などを今後明らかにする。
Studies on the crystal structure of antiporters for organic compounds and the mechanisms
Grant number：13142205 2001 - 2005
Japan Society for the Promotion of Science Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research on Priority Areas
YAMAGUCHI Akihito, MURAKAMI Satoshi, HIRATA Takahiro
Grant amount：\135400000 （ Direct Cost: \135400000 ）
This project aimed for determination of the crystal structure of bacterial major xenobiotic exporter AcrB and to reveal the mechanism of xenobiotic recognition and efflux transport. Before this project, no crystal structures of membrane transporters had determined at all. Because the purpose of this project was highly riskful, at first we had one more target to determine two-dimensional structure of AcrB by using electron microscopic analysis as a risk hedge. However, as a result, the risk hedge was not required.
Murakami et al. in our laboratory succeeded to determine the crystal structure of AcrB one year after starting this project. It is not only the first structure of xenobiotic exporters but also the first structure of the secondary transporters. This achievement is highly evaluated as a milestone of the membrane transport study. On the basis of this crystal structure, the xenobiotic recognition mechanism is clearly revealed: The AcrB structure has its entrance for substrates at the side of the molecule opened to the lipid bilayer region of the membrane. AcrB acts as a membrane vacuum cleaner by taking up its substrates from the lipid bilayer region. Because xenobiotics generally enter into cells through lipid bilayer region by simple diffusion, this mechanism efficiently recognize xenobiotics.
Our first structure did not contain bound substrate, therefore, it can not answer the question, "where is the binding site?". On the basis of the first structure, it was estimated as the central cavity of the trimer. One years after, Edward Yu et al. reported that the structure of AcrB in which substrates bound at the central cavity. However, this estimation was no consisted with the mutational studies; there is no amino acid residues important for substrate recognition at the central cavity.
In the last year of this project, Murakami's group succeeded to determine the substrate binding structure of AcrB. The trimer is asymmetric. Only one protomer binds substrate at the phenylalanine cluster region, which is different from the central cavity. The other two porotmers represent the substrate extrusion step and the standing for substrate access step, respectively. In other words, this one crystal contains all snapshots for three steps of efflux transport. On the basis of this crystal structure, it was revealed that substrates are transported by the completely new mechanism named as functionally rotating binding change mechanism similar to FoF1-ATPase. In addition, it was revealed that the extraordinary broad substrate recognition is achieved by the multisite binding with mainly hydrophobic interactions.
Post-Genomic Approach to Bacterial Xenobiotic Exporter Gene Resources and Investigation of Novel Drug Resistance Mechanisms of Bacteria
Grant number：13854012 2001 - 2005
Japan Society for the Promotion of Science Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (S)
YAMAGUCHI Akihito, MURAKAMI Satoshi, HIRATA Takahiro
Grant amount：\123370000 （ Direct Cost: \94900000 、 Indirect Cost：\28470000 ）
This project aimed to investigate comprehensive analysis of bacterial xenobiotic exporters including their expression regulation mechanisms using mainly Escherichia coli as a model. We previously established the complete expression-cloning library of E.coli putative xenobiotic exporters. On the basis of the library, we analyzed the four typical groups of xenobiotic exporters. Among them, we found that all five of the RND exporters, two of the MFS transporters and one of the ABC transporters couple with the same outer membrane channel TolC. Before that, MFS and ABC exporters had not estimated to have large periplasmic domain required for TolC interaction from their sequence analysis. So, we determined the topology of MacB (ABC) and EmrB (MFS) by site-directed chemical modification and revealed that MacB and EmrB have only four and five transmembrane helices, respectively, and contain large periplasmic loops enough to interact with TolC.
As for the major xenobiotic exporter AcrB (RND) in E.coli, we succeeded to determine the X-ray crystallographic structure in 2002, which was the first structure not only as xenobiotic exporters but also as secondary transporters. Then, last year, we succeeded to elucidate the structure of the substrate-binding form of AcrB. As a result, we revealed that xenobiotic recognition is based on the membrane vacuum cleaner mechanism from outer leaflet of the inner membrane.
As for the expression control of xenobiotic exporter genes, we first found that two-component signal transduction systems, which are the bacterial environmental response systems, induce xenobiotic exporter expression. We revealed the complex network of xenobiotic exporter gene expression by two-component systems. Besides, we found that indole acts as an intercellular signal transduction molecule and induce some xenobiotic exporter genes. Xenobiotic exporters were also controlled by the regulatory systems for ferrous homeostasis.
In addition, we found that xenobiotic exporters confer not only to xenobiotic extrusion but also bacterial pathogenicity. That is, although mice which were perorally inoculated Salmonella typhimurium died within a few days, peroral inoculation of S.typhimurium of which all major xenobiotic exporter genes were knocked out did not cause death. Among them, the knock out of MacB gene, which is the only macrolide-specific exporter, resulted in almost nonpoisonous bacteria. These results strongly suggested that some xenobiotic exporters must have their intrinsic physiological roles other than xenobiotic export. These findings suggest the possibility of a completely novel antibacterial drug target which removes pathogenicity without killing bacteria. Coexistence with bacteria is expected to greatly decrease the emergence of drug resistant pathogens.
テトラサイクリン排出蛋白の結晶構造解析
Grant number：11878112 1999
日本学術振興会 科学研究費助成事業 萌芽的研究
山口 明人, 村上 聡
Grant amount：\2100000 （ Direct Cost: \2100000 ）
1、テトラサイクリン排出タンパクの結晶化及び結晶構造解析に向けて、その基盤となる大量産生と大量精製法の条件検討を行った。その結果、1週間ほどの培養〜精製の行程で、30mg程の純化したテトラサイクリン排出タンパクを得る系の確立に成功した。この量は結晶化の実験に用いるには不足は無く、結晶化実験に向け基盤を固めることができた。
2、膜蛋白質の結晶化に於いて最も影響の大きな因子の一つである界面活性剤の選択に関して本年度は、その可溶化力を市販の物ほぼ全てに関してその効果を検討した。約100種類の界面活性剤のうち、ある種の合成リン脂質誘導体が蛋白質を効率よく膜から可溶化し、且つ精製後も蛋白質標品を安定に保つ事を見出した。この界面活性剤で安定化させたテトラサイクリン排出タンパクは水溶液中でも安定で結晶化が起こる一般的な蛋白濃度である10mg/mLもの高濃度でも非特異的会合を起こさない事が分かった。今後、界面活性剤の蛋白質標品の安定化に及ぼす影響をさらに検討する。
3、生物分子工学研究所(BERI)木村能章博士との共同研究により、精製したテトラサイクリン排出タンパク分子を合成リン脂質シートに再構成し、相転移によりシート中のたんぱく質濃度を上昇させる手法で初歩的な2次元結晶化に成功した。このシートの電顕像のフーリエ変換像には分子配列に規則性があることを示すピークが認められた。
4、3次元結晶化に関しては、市販の結晶化条件スクリーニングキットを用いて初歩的なスクリーニングを行った。今後リン脂質の系に再構成する条件検討の結果を応用し、Lipid cubic phase 法による結晶化を行って行く予定である。
Molecular Basis and Physiological Roles of Xenobiotic Exporters
Grant number：10308029 1998 - 2000
Japan Society for the Promotion of Science Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (A).
YAMAGUCHI Akihito, NADA Shigeyuki, MURAKAMI Satoshi
Grant amount：\31800000 （ Direct Cost: \31800000 ）
(1) We constructed the complete set of cysteine-scanning mutants of tetracycline/H+ antiporter TetA (B) and investigated the detailed topology by means of site-directed chemical modification and oxidative disulfide cross-linking formation of double Cys mutants. As a result, it was revealed that TetA (B) forms a circular form and involves a water-filled channel at the center of the molecule with a permeability barrier in the middle of the membrane.
(2) We established the method for measuring the drug efflux mediated by MRP2 by means of fluorescent drugs. Then, we determined the transmembrane basic residues essential for its transport function and the binding of the inhibitor by means of site-directed mutagenesis.
(3) We first succeeded to obtain crystals of xenobiotic exporters. Now, we are undergoing the X-ray crystallographic analysis using SPring8.
(4) We found a novel possible exporter for sphingosine-1-phosphate from platelete and its complete cDNA was cloned and expressed in a cell line.
