Research activity information

• Jun. 2011 一般社団法人神緑会, 田中千賀子学術奨励賞, 女性研究者

  Shima Fumi

• SARS-CoV-2 papain-like protease inhibits ISGylation of the viral nucleocapsid protein to evade host anti-viral immunity

  Aulia Fitri Rhamadianti, Takayuki Abe, Tomohisa Tanaka, Chikako Ono, Hisashi Katayama, Yoshiteru Makino, Lin Deng, Chieko Matsui, Kohji Moriishi, Fumi Shima, Yoshiharu Matsuura, Ikuo Shoji

  ABSTRACT A severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection causes mild-to-severe respiratory symptoms, including acute respiratory distress. Despite remarkable efforts to investigate the virological and pathological impacts of SARS-CoV-2, many of the characteristics of SARS-CoV-2 infection still remain unknown. The interferon-inducible ubiquitin-like protein ISG15 is covalently conjugated to several viral proteins to suppress their functions. It was reported that SARS-CoV-2 utilizes its papain-like protease (PLpro) to impede ISG15 conjugation, ISGylation. However, the role of ISGylation in SARS-CoV-2 infection remains unclear. We aimed to elucidate the role of ISGylation in SARS-CoV-2 replication. We observed that the SARS-CoV-2 nucleocapsid protein is a target protein for the HERC5 E3 ligase-mediated ISGylation in cultured cells. Site-directed mutagenesis reveals that the residue K374 within the C-terminal spacer B-N3 (SB/N3) domain is required for nucleocapsid-ISGylation, alongside conserved lysine residue in MERS-CoV (K372) and SARS-CoV (K375). We also observed that the nucleocapsid-ISGylation results in the disruption of nucleocapsid oligomerization, thereby inhibiting viral replication. Knockdown of ISG15 mRNA enhanced SARS-CoV-2 replication in the SARS-CoV-2 reporter replicon cells, while exogenous expression of ISGylation components partially hampered SARS-CoV-2 replication. Taken together, these results suggest that SARS-CoV-2 PLpro inhibits ISGylation of the nucleocapsid protein to promote viral replication by evading ISGylation-mediated disruption of the nucleocapsid oligomerization. IMPORTANCE ISG15 is an interferon-inducible ubiquitin-like protein that is covalently conjugated to the viral protein via specific Lys residues and suppresses viral functions and viral propagation in many viruses. However, the role of ISGylation in SARS-CoV-2 infection remains largely unclear. Here, we demonstrated that the SARS-CoV-2 nucleocapsid protein is a target protein for the HERC5 E3 ligase-mediated ISGylation. We also found that the residue K374 within the C-terminal spacer B-N3 (SB/N3) domain is required for nucleocapsid-ISGylation. We obtained evidence suggesting that nucleocapsid-ISGylation results in the disruption of nucleocapsid-oligomerization, thereby suppressing SARS-CoV-2 replication. We discovered that SARS-CoV-2 papain-like protease inhibits ISG15 conjugation of nucleocapsid protein via its de-conjugating enzyme activity. The present study may contribute to gaining new insight into the roles of ISGylation-mediated anti-viral function in SARS-CoV-2 infection and may lead to the development of more potent and selective inhibitors targeted to SARS-CoV-2 nucleocapsid protein.

  American Society for Microbiology, Sep. 2024, Journal of Virology, 98(9) (9), English

  Scientific journal

  Link to Kobe Univ. Repository (Kernel)


• Small-molecule RAS/RAF Binding Inhibitors Allosterically Disrupt RAF Conformation and Exert Efficacy Against a Broad Spectrum of RAS-driven Cancers

  Fumi Shima, Yoko Yoshikawa, Yoshiteru Makino, Hirokazu Kubota, Takashi Kawamura, Shigeyuki Matsumoto, Hitomi Yuki, Akira Shibaike, Megumi Okamura, Tomoyo Okada, Manabu Horikawa, Kazumasa Horie, Michiyo Koyanagi-Aoi, Takashi Aoi, Tohru Kataoka, Teruki Honma, Takashi Kumasaka, Hiroo Koyama

  Jan. 2024


• Oncogenic mutations Q61L and Q61H confer active form-like structural features to the inactive state (state 1) conformation of H-Ras protein

  Shigeyuki Matsumoto, Haruka Taniguchi-Tamura, Mitsugu Araki, Takashi Kawamura, Ryo Miyamoto, Chiemi Tsuda, Fumi Shima, Takashi Kumasaka, Yasushi Okuno, Tohru Kataoka

  GTP-bound forms of Ras proteins (Ras•GTP) assume two interconverting conformations, "inactive" state 1 and "active" state 2. Our previous study on the crystal structure of the state 1 conformation of H-Ras in complex with guanosine 5'-(β, γ-imido)triphosphate (GppNHp) indicated that state 1 is stabilized by intramolecular hydrogen-bonding interactions formed by Gln61. Since Ras are constitutively activated by substitution mutations of Gln61, here we determine crystal structures of the state 1 conformation of H-Ras•GppNHp carrying representative mutations Q61L and Q61H to observe the effect of the mutations. The results show that these mutations alter the mode of hydrogen-bonding interactions of the residue 61 with Switch II residues and induce conformational destabilization of the neighboring regions. In particular, Q61L mutation results in acquirement of state 2-like structural features. Moreover, the mutations are likely to impair an intramolecular structural communication between Switch I and Switch II. Molecular dynamics simulations starting from these structures support the above observations. These findings may give a new insight into the molecular mechanism underlying the aberrant activation of the Gln61 mutants.

  Elsevier BV, Aug. 2021, Biochemical and Biophysical Research Communications, 565, 85 - 90, English, International magazine

  [Refereed]

  Scientific journal

  Link to Kobe Univ. Repository (Kernel)


• Molecular Basis for Allosteric Inhibition of GTP-Bound H-Ras Protein by a Small-Molecule Compound Carrying a Naphthalene Ring.

  Matsumoto S, Hiraga T, Hayashi Y, Yoshikawa Yoko, Tsuda C, Araki M, Neya M, SHIMA FUMI, Kataoka Tohru

  Sep. 2018, Biochemistry, 57(36) (36), 5350 - 5358, English

  [Refereed]

  Scientific journal

  Link to Kobe Univ. Repository (Kernel)


• Ras inhibitors display an anti-metastatic effect by downregulation of lysyl oxidase through inhibition of the Ras-PI3K-Akt-HIF-1 alpha pathway

  Yoko Yoshikawa, Osamu Takano, Ichiro Kato, Yoshihisa Takahashi, Fumi Shima, Tohru Kataoka

  Metastasis stands as the major obstacle for the survival from cancers. Nonetheless most existing anti-cancer drugs inhibit only cell proliferation, and discovery of agents having both anti-proliferative and anti-metastatic properties would be more beneficial. We previously reported the discovery of small-molecule Ras inhibitors, represented by Kobe0065, that displayed anti-proliferative activity on xenografts of human colorectal cancer (CRC) cell line SW480 carrying the K-ras(G12V) gene. Here we show that treatment of cancer cells carrying the activated ras genes with Kobe0065 or a siRNA targeting Ras downregulates the expression of lysyl oxidase (LOX), which has been implicated in metastasis. LOX expression is enhanced by co-expression of Ras(G12V) through activation of phosphatidylinositol 3-kinase (PI3K)/Akt and concomitant accumulation of hypoxia-inducible factor (HIF)-1 alpha. Furthermore, Kobe0065 effectively inhibits not only migration and invasion of cancer cells carrying the activated ras genes but also lung metastasis of human CRC cell line SW620 carrying the K-ras(G12V) gene. Collectively, these results indicate that Kobe0065 prevents metastasis through inhibition of the Ras-P13K-Akt-HIF-1 alpha-LOX signaling and suggest that Ras inhibitors in general might exhibit both anti-proliferative and anti-metastatic properties toward cancer cells carrying the activated ras genes. (C) 2017 Elsevier B.V. All rights reserved.

  ELSEVIER IRELAND LTD, Dec. 2017, CANCER LETTERS, 410, 82 - 91, English

  [Refereed]

  Scientific journal

  Link to Kobe Univ. Repository (Kernel)


• Structural transition of solvated H-Ras/GTP revealed by molecular dynamics simulation and local network entropy

  Shota Matsunaga, Yuta Hano, Yuta Saito, Kazuhiro J. Fujimoto, Takashi Kumasaka, Shigeyuki Matsumoto, Tohru Kataoka, Fumi Shima, Shigenori Tanaka

  The state transitions of solvated H-Ras protein with GTP were theoretically analyzed through molecular dynamics (MD) simulations. To accelerate the structural changes associated with the locations of two switch regions (I and II), the Parallel Cascade Selection MD (PaCS-MD) method was employed in this study. The interconversions between the State 1 and State 2 were thus studied in atomic details, leading to a reasonable agreement with experimental observations and consequent scenarios concerning the transition mechanism that would be essential for the development of Ras inhibitors as anti-cancer agents. Furthermore, the state-transition-based local network entropy (SNE) was calculated for the transition process from State 1 to State 2, by which the temporal evolution of information entropy associated with the dynamical behavior of hydrogen bond network composed of hydration water molecules was described. The calculated results of SNE thus proved to provide a good indicator to detect the dynamical state transition of solvated Ras protein system (and probably more general systems) from a viewpoint of nonequilibrium statistical thermodynamics. (C) 2017 Elsevier Inc. All rights reserved.

  ELSEVIER SCIENCE INC, Oct. 2017, JOURNAL OF MOLECULAR GRAPHICS & MODELLING, 77, 51 - 63, English

  [Refereed]

  Scientific journal

  Link to Kobe Univ. Repository (Kernel)


• Integrin-alpha 10 Dependency Identifies RAC and RICTOR as Therapeutic Targets in High-Grade Myxofibrosarcoma

  Tomoyo Okada, Ann Y. Lee, Li-Xuan Qin, Narasimhan Agaram, Takahiro Mimae, Yawei Shen, Rachael O'Connor, Miguel A. Lopez-Lago, Amanda Craig, Martin L. Miller, Phaedra Agius, Evan Molinelli, Nicholas D. Socci, Aimee M. Crago, Fumi Shima, Chris Sander, Samuel Singer

  Myxofibrosarcoma is a common mesenchymal malignancy with complex genomics and heterogeneous clinical outcomes. Through gene-expression profiling of 64 primary high-grade myxofibrosarcomas, we defined an expression signature associated with clinical outcome. The gene most significantly associated with disease-specific death and distant metastasis was ITGA10 (integrin-alpha 10). Functional studies revealed that myxofibrosarcoma cells strongly depended on integrin-alpha 10, whereas normal mesenchymal cells did not. Integrin-alpha 10 transmitted its tumor-specific signal via TRIO and RICTOR, two oncoproteins that are frequently co-overexpressed through gene amplification on chromosome 5p. TRIO and RICTOR activated RAC/PAK and AKT/mTOR to promote sarcoma cell survival. Inhibition of these proteins with EHop-016 (RAC inhibitor) and INK128 (mTOR inhibitor) had antitumor effects in tumor-derived cell lines and mouse xenografts, and combining the drugs enhanced the effects. Our results demonstrate the importance of integrin-alpha 10/TRIO/RICTOR signaling for driving myxofibrosarcoma progression and provide the basis for promising targeted treatment strategies for patients with high-risk disease. SIGNIFICANCE: Identifying the molecular pathogenesis for myxofibrosarcoma progression has proven challenging given the highly complex genomic alterations in this tumor type. We found that integrin-alpha 10 promotes tumor cell survival through activation of TRIO-RAC-RICTOR-mTOR signaling, and that inhibitors of RAC and mTOR have antitumor effects in vivo, thus identifying a potential treatment strategy for patients with high-risk myxofibrosarcoma. (C) 2016 AACR.

  AMER ASSOC CANCER RESEARCH, Oct. 2016, CANCER DISCOVERY, 6(10) (10), 1148 - 1165, English

  [Refereed]

  Scientific journal


• Molecular Mechanism for Conformational Dynamics of Ras.GTP Elucidated from In-Situ Structural Transition in Crystal

  Shigeyuki Matsumoto, Nao Miyano, Seiki Baba, Jingling Liao, Takashi Kawamura, Chiemi Tsuda, Azusa Takeda, Masaki Yamamoto, Takashi Kumasaka, Tohru Kataoka, Fumi Shima

  Ras.GTP adopts two interconverting conformational states, state 1 and state 2, corresponding to inactive and active forms, respectively. However, analysis of the mechanism for state transition was hampered by the lack of the structural information on wild-type Ras state 1 despite its fundamental nature conserved in the Ras superfamily. Here we solve two new crystal structures of wild-type H-Ras, corresponding to state 1 and state 2. The state 2 structure seems to represent an intermediate of state transition and, intriguingly, the state 1 crystal is successfully derived from this state 2 crystal by regulating the surrounding humidity. Structural comparison enables us to infer the molecular mechanism for state transition, during which a wide range of hydrogen-bonding networks across Switch I, Switch II and the alpha 3-helix interdependently undergo gross rearrangements, where fluctuation of Tyr32, translocation of Gln61, loss of the functional water molecules and positional shift of GTP play major roles. The NMR-based hydrogen/deuterium exchange experiments also support this transition mechanism. Moreover, the unveiled structural features together with the results of the biochemical study provide a new insight into the physiological role of state 1 as a stable pool of Ras.GTP in the GDP/GTP cycle of Ras.

  NATURE PUBLISHING GROUP, May 2016, SCIENTIFIC REPORTS, 6, 25931, English

  [Refereed]

  Scientific journal

  Link to Kobe Univ. Repository (Kernel)


• In silico discovery of small-molecule Ras inhibitors that display antitumor activity by blocking the Ras-effector interaction

  Fumi Shima, Yoko Yoshikawa, Min Ye, Mitsugu Araki, Shigeyuki Matsumoto, Jingling Liao, Lizhi Hu, Takeshi Sugimoto, Yuichi Ijiri, Azusa Takeda, Yuko Nishiyama, Chie Sato, Shin Muraoka, Atsuo Tamura, Tsutomu Osoda, Ken-ichiro Tsuda, Tomoya Miyakawa, Hiroaki Fukunishi, Jiro Shimada, Takashi Kumasaka, Masaki Yamamoto, Tohru Kataoka

  Mutational activation of the Ras oncogene products (H-Ras, K-Ras, and N-Ras) is frequently observed in human cancers, making them promising anticancer drug targets. Nonetheless, no effective strategy has been available for the development of Ras inhibitors, partly owing to the absence of well-defined surface pockets suitable for drug binding. Only recently, such pockets have been found in the crystal structures of a unique conformation of Ras center dot GTP. Here we report the successful development of small-molecule Ras inhibitors by an in silico screen targeting a pocket found in the crystal structure of M-Ras center dot GTP carrying an H-Ras-type substitution P40D. The selected compound Kobe0065 and its analog Kobe2602 exhibit inhibitory activity toward H-Ras center dot GTP-c-Raf-1 binding both in vivo and in vitro. They effectively inhibit both anchorage-dependent and -independent growth and induce apoptosis of H-ras(G12V)-transformed NIH 3T3 cells, which is accompanied by down-regulation of downstream molecules such as MEK/ERK, Akt, and RalA as well as an upstream molecule, Son of sevenless. Moreover, they exhibit antitumor activity on a xenograft of human colon carcinoma SW480 cells carrying the K-ras(G12V) gene by oral administration. The NMR structure of a complex of the compound with H-Ras center dot GTP(T35S), exclusively adopting the unique conformation, confirms its insertion into one of the surface pockets and provides a molecular basis for binding inhibition toward multiple Ras center dot GTP-interacting molecules. This study proves the effectiveness of our strategy for structure-based drug design to target Ras center dot GTP, and the resulting Kobe0065-family compounds may serve as a scaffold for the development of Ras inhibitors with higher potency and specificity.

  NATL ACAD SCIENCES, May 2013, PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 110(20) (20), 8182 - 8187, English

  [Refereed]

  Scientific journal


• Discovery of Small-Molecule Ras Inhibitors that Display Antitumor Activity by Interfering with Ras·GTP-Effector Interaction

  Fumi Shima, Yoko Yoshikawa, Shigeyuki Matsumoto, Tohru Kataoka

  Ras proteins, particularly their active GTP-bound forms (Ras·GTP), were thought "undruggable" owing to the absence of apparent drug-accepting pockets in their crystal structures. Only recently, such pockets have been found in the crystal structures representing a novel Ras·GTP conformation. We have conducted an in silico docking screen targeting a pocket in the crystal structure of M-RasP40D·GTP and obtained Kobe0065, which, along with its analogue Kobe2602, inhibits binding of H-Ras·GTP to c-Raf-1. They inhibit the growth of H-rasG12V-transformed NIH3T3 cells, which are accompanied by downregulation of not only MEK/ERK but also Akt, RalA, and Sos, indicating the blockade of interaction with multiple effectors. Moreover, they exhibit antitumor activity on a xenograft of human colon carcinoma carrying K-rasG12V. The nuclear magnetic resonance structure of a complex of the compound with H-RasT35S·GTP confirms its insertion into the surface pocket. Thus, these compounds may serve as a novel scaffold for the development of Ras inhibitors with higher potency and specificity. © 2013 Elsevier Inc.

  2013, Enzymes, 34, 1 - 23, English

  [Refereed]

  Scientific journal


• Structure-based drug design of small-molecule Ras inhibitors having anti-tumor activity

  Shima Fumi, Kataoka T

  2013, SPring-8 Research Frontiers, English

  [Refereed][Invited]

  Research institution


• Crystal structures of the state 1 conformations of the GTP-bound H-Ras protein and its oncogenic G12V and Q61L mutants

  Shin Muraoka, Fumi Shima, Mitsugu Araki, Tomoko Inoue, Akiko Yoshimoto, Yuichi Ijiri, Nobuaki Seki, Atsuo Tamura, Takashi Kumasaka, Masaki Yamamoto, Tohru Kataoka

  GTP-bound Ras adopts two interconverting conformations, "inactive" state 1 and "active" state 2. However, the tertiary structure of wild-type (WT) state 1 remains unsolved. Here we solve the state 1 crystal structures of H-Ras WT together with its oncogenic G12V and Q61L mutants. They assume open structures characterized by impaired interactions of both Thr-35 in switch I and Gly-60 in switch II with the gamma-phosphate of GTP and possess two surface pockets of mutually different shapes unseen in state 2, a potential target for selective inhibitor development. Furthermore, they provide a structural basis for the low GTPase activity of state 1. (c) 2012 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

  ELSEVIER SCIENCE BV, Jun. 2012, FEBS LETTERS, 586(12) (12), 1715 - 1718, English

  [Refereed]

  Scientific journal


• Solution Structure of the State 1 Conformer of GTP-bound H-Ras Protein and Distinct Dynamic Properties between the State 1 and State 2 Conformers

  Mitsugu Araki, Fumi Shima, Yoko Yoshikawa, Shin Muraoka, Yuichi Ijiri, Yuka Nagahara, Tomoya Shirono, Tohru Kataoka, Atsuo Tamura

  Ras small GTPases undergo dynamic equilibrium of two interconverting conformations, state 1 and state 2, in the GTP-bound forms, where state 2 is recognized by effectors, whereas physiological functions of state 1 have been unknown. Limited information, such as static crystal structures and (31)P NMR spectra, was available for the study of the conformational dynamics. Here we determine the solution structure and dynamics of state 1 by multidimensional heteronuclear NMR analysis of an H-RasT35S mutant in complex with guanosine 5'-(beta, gamma-imido)triphosphate (GppNHp). The state 1 structure shows that the switch I loop fluctuates extensively compared with that in state 2 or H-Ras-GDP. Also, backbone (1)H, (15)N signals for state 2 are identified, and their dynamics are studied by utilizing a complex with c-Raf-1. Furthermore, the signals for almost all the residues of H-Ras.GppNHp are identified by measurement at low temperature, and the signals for multiple residues are found split into two peaks corresponding to the signals for state 1 and state 2. Intriguingly, these residues are located not only in the switch regions and their neighbors but also in the rigidly structured regions, suggesting that global structural rearrangements occur during the state interconversion. The backbone dynamics of each state show that the switch loops in state 1 are dynamically mobile on the picosecond to nanosecond time scale, and these mobilities are significantly reduced in state 2. These results suggest that multiconformations existing in state 1 are mostly deselected upon the transition toward state 2 induced by the effector binding.

  AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC, Nov. 2011, JOURNAL OF BIOLOGICAL CHEMISTRY, 286(45) (45), 39644 - 39653, English

  [Refereed]

  Scientific journal


• Critical Roles of Interactions among Switch I-preceding Residues and between Switch II and Its Neighboring alpha-Helix in Conformational Dynamics of the GTP-bound Ras Family Small GTPases

  Kousuke Matsumoto, Fumi Shima, Shin Muraoka, Mitsugu Araki, Lizhi Hu, Yuichi Ijiri, Rina Hirai, Jingling Liao, Takashi Yoshioka, Takashi Kumasaka, Masaki Yamamoto, Atsuo Tamura, Tohru Kataoka

  GTP-bound forms of Ras family small GTPases exhibit dynamic equilibrium between two interconverting conformations, "inactive" state 1 and "active" state 2. A great variation exists in their state distribution; H-Ras mainly adopts state 2, whereas M-Ras predominantly adopts state 1. Our previous studies based on comparison of crystal structures representing state 1 and state 2 revealed the importance of the hydrogen-bonding interactions of two flexible effector-interacting regions, switch I and switch II, with the gamma-phosphate of GTP in establishing state 2 conformation. However, failure to obtain both state structures from a single protein hampered further analysis of state transition mechanisms. Here, we succeed in solving two crystal structures corresponding to state 1 and state 2 from a single Ras polypeptide, M-RasD41E, carrying an H-Ras-type substitution in residue 41, immediately preceding switch I, in complex with guanosine 5'-(beta,gamma-imido)triphosphate. Comparison among the two structures and other state 1 and state 2 structures of H-Ras/M-Ras reveal two new structural features playing critical roles in state dynamics; interaction of residues 31/41 (H-Ras/M-Ras) with residues 29/39 and 30/40, which induces a conformational change of switch I favoring its interaction with the gamma-phosphate, and the hydrogen-bonding interaction of switch II with its neighboring alpha-helix, alpha 3-helix, which induces a conformational change of switch II favoring its interaction with the gamma-phosphate. The importance of the latter interaction is proved by mutational analyses of the residues involved in hydrogen bonding. These results define the two novel functional regions playing critical roles during state transition.

  AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC, Apr. 2011, JOURNAL OF BIOLOGICAL CHEMISTRY, 286(17) (17), 15403 - 15412, English

  [Refereed]

  Scientific journal


• Structural Basis for Conformational Dynamics of GTP-bound Ras Protein

  Fumi Shima, Yuichi Ijiri, Shin Muraoka, Jingling Liao, Min Ye, Mitsugu Araki, Kousuke Matsumoto, Naoki Yamamoto, Takeshi Sugimoto, Yoko Yoshikawa, Takashi Kumasaka, Masaki Yamamoto, Atsuo Tamura, Tohru Kataoka

  Ras family small GTPases assume two interconverting conformations, "inactive" state 1 and "active" state 2, in their GTP-bound forms. Here, to clarify the mechanism of state transition, we have carried out x-ray crystal structure analyses of a series of mutant H-Ras and M-Ras in complex with guanosine 5'-(beta,gamma-imido) triphosphate (GppNHp), representing various intermediate states of the transition. Crystallization of H-RasT35S-GppNHp enables us to solve the first complete tertiary structure of H-Ras state 1 possessing two surface pockets unseen in the state 2 or H-Ras-GDP structure. Moreover, determination of the two distinct crystal structures of H-RasT35S-GppNHp, showing prominent polysterism in the switch I and switch II regions, reveals a pivotal role of the guanine nucleotide-mediated interaction between the two switch regions and its rearrangement by a nucleotide positional change in the state 2 to state 1 transition. Furthermore, the (31)P NMR spectra and crystal structures of the GppNHp-bound forms of M-Ras mutants, carrying various H-Ras-type amino acid substitutions, also reveal the existence of a surface pocket in state 1 and support a similar mechanism based on the nucleotide-mediated interaction and its rearrangement in the state 1 to state 2 transition. Intriguingly, the conformational changes accompanying the state transition mimic those that occurred upon GDP/GTP exchange, indicating a common mechanistic basis inherent in the high flexibility of the switch regions. Collectively, these results clarify the structural features distinguishing the two states and provide new insights into the molecular basis for the state transition of Ras protein.

  AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC, Jul. 2010, JOURNAL OF BIOLOGICAL CHEMISTRY, 285(29) (29), 22696 - 22705, English

  [Refereed]

  Scientific journal


• Biochemical characterization of missense mutations in the Arf/Arl-family small GTPase Arl6 causing Bardet-Biedl syndrome

  Tetsuo Kobayashi, Yuji Hori, Nami Ueda, Hiroaki Kajiho, Shin Muraoka, Fumi Shima, Tohru Kataoka, Kenji Kontani, Toshiaki Katada

  Bardet-Biedl syndrome (BBS) is a pleiotropically genetic disorder, whose etiology is linked to cilia. Mutations in the Arf/Arl-family GTPase Arl6 have been recently shown to be responsible for BBS type 3. Here we show that BBS mutations alter the guanine nucleotide-binding properties of Arl6. Specifically, substitution of 31st Threonine to Arginine selectively abrogates the GTP-binding ability of Arl6 without affecting GDP binding/dissociating properties. Furthermore, all the BBS Mutations in Arl6 result in low expression of the mutant proteins, Which can be restored by the inhibition of the proteasome. These findings implicate that Arl6 Mutants are destabilized and eliminated by the proteasome in cells, probably due to the altered nucleotide-binding properties. (C) 2009 Elsevier Inc. All rights reserved.

  ACADEMIC PRESS INC ELSEVIER SCIENCE, Apr. 2009, BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 381(3) (3), 439 - 442, English

  [Refereed]

  Scientific journal


• Two conformational states of Ras GTPase exhibit differential GTP-binding kinetics

  Jingling Liao, Fumi Shima, Mitsugu Araki, Min Ye, Shin Muraoka, Takeshi Sugimoto, Mei Kawamura, Naoki Yamamoto, Atsuo Tamura, Tohru Kataoka

  Previous P-31 NMR studies revealed that small GTPases H-Ras and K-Ras in complex with GTP assume two interconverting conformational states, state I and state 2. While state 2 corresponds to an active conformation, little is known about the function of state 1, an inactive conformation incapable of effector binding. To address the biochemical properties of state 1, we measured the P-31 NMR spectra of five Ras family small GTPases; H-Ras, M-Ras, Rap1A, Rap2A and Ra1A, and find that they exhibit distinctive state 2/state 1 populations with the ratios ranging from 0.072 for M-Ras to 16 for Rap2A. Further, we show that GTPases with higher populations of state I exhibit higher dissociation and association rate constants for GTP. These results imply that GTP loading to the nucleotide-free small GTPases preferentially yields state 1, which is subsequently converted to state 2, rendering the GTP-bound form functional. (c) 2008 Elsevier Inc. All rights reserved.

  ACADEMIC PRESS INC ELSEVIER SCIENCE, May 2008, BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 369(2) (2), 327 - 332, English

  [Refereed]

  Scientific journal


• Crystal structure of M-Ras reveals a GTP-bound "off" state conformation of Ras family small GTPases

  M Ye, F Shima, S Muraoka, JL Liao, H Okamoto, M Yamamoto, A Tamura, N Yagi, T Ueki, T Kataoka

  Although some members of Ras family small GTPases, including M-Ras, share the primary structure of their effector regions with Ras, they exhibit vastly different binding properties to Ras effectors such as c-Raf-1. We have solved the crystal structure of M-Ras in the GDP-bound and guanosine 5'-(beta, gamma-imido) triphosphate (Gpp(NH) p)-bound forms. The overall structure of M-Ras resembles those of H-Ras and Rap2A, except that M-Ras-Gpp( NH) p exhibits a distinctive switch I conformation, which is caused by impaired intramolecular interactions between Thr-45 (corresponding to Thr-35 of H-Ras) of the effector region and the gamma-phosphate of Gpp(NH) p. Previous P-31 NMR studies showed that H-Ras-Gpp( NH) p exists in two interconverting conformations, states 1 and 2. Whereas state 2 is a predominant form of H-Ras and corresponds to the "on" conformation found in the complex with effectors, state 1 is thought to represent the "off" conformation, whose tertiary structure remains unknown. P-31 NMR analysis shows that free M-Ras-Gpp(NH)p predominantly assumes the state 1 conformation, which undergoes conformational transition to state 2 upon association with c-Raf-1. These results indicate that the solved structure of M-Ras-Gpp(NH) p corresponds to the state 1 conformation. The predominance of state 1 in M-Ras is likely to account for its weak binding ability to the Ras effectors, suggesting the importance of the tertiary structure factor in small GTPase-effector interaction. Further, the first determination of the state 1 structure provides a molecular basis for developing novel anti-cancer drugs as compounds that hold Ras in the state 1 "off" conformation.

  AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC, Sep. 2005, JOURNAL OF BIOLOGICAL CHEMISTRY, 280(35) (35), 31267 - 31275, English

  [Refereed]

  Scientific journal


• Critical role of posttranslational modification of Ras proteins in effector activation

  F Shima, T Kataoka

  JAPANESE BIOCHEMICAL SOC, Jun. 2005, SEIKAGAKU, 77(6) (6), 519 - 526, Japanese

  [Refereed]

  Scientific journal


• Direct activation of fission yeast adenylyl cyclase by heterotrimeric G protein gpa2

  Hideki Ogihara, Fumi Shima, Kana Naito, Tsuyoshi Asato, Ken-Ichi Kariya, Tohru Kataoka

  Genetic studies on Schizosaccharomyces pombe adenylyl cyclase (cyr1) have shown that its activity is positively regulated by a heterotrimetric G protein α subunit gpa2 and that the resulting increase in intracellular cAMP concentration causes inhibition of sexual development including mating and meiosis. However, molecular mechanism underlying this gpa2-dependent regulation of cyr1 remains to be clarified. Here, we show that gpa2 exhibits a direct and GTP-dependent binding to the Ras-associating domain (RAD) of cyr1, which is identified by a computer algorithm-based search of the cyr1 amino acid sequence. Overexpression of this RAD results in acceleration of the sexual development of fission yeast cells presumably by competitive sequestration of gpa2. Furthermore, cyr1 is activated in vitro by the addition of purified gpa2, which is converted to the active state by treatment with AlF4-. These results indicate a crucial role of the RAD as a direct binding site of gpa2 in activation of cyr1. Thus, RADs, which have been defined as a conserved motif shared among the Ras-family small G protein-associating domains, are for the first time shown to exhibit a functional association with a member of the heterotrimeric G proteins.

  2004, Kobe Journal of Medical Sciences, 50(3-4) (3-4), 111 - 121, English

  [Refereed]

  Scientific journal

  Link to Kobe Univ. Repository (Kernel)


• Critical function of the ras-associating domain as a primary ras-binding site for regulation of Saccharomyces cerevisiae adenylyl cyclase

  M Kido, F Shima, T Satoh, T Asato, K Kariya, T Kataoka

  Mammalian candidate effectors of the small GTPase Ras, such as RalGDS, afadin/AF-6, Rin1, and phospholipase Cc, have been shown to share structurally conserved modules termed Ras-associating (RA) domains at their Ras-binding sites. The Ras-binding domains of Raf-1 and phosphoinositide 3-kinase gamma (other Ras effectors) also share a similar tertiary structure with the RA domains. On the other hand, the primary Ras-binding site of Saccharomyces cerevisiae adenylyl cyclase, the best characterized Ras effector, has been mapped by mutational studies to the leucine-rich repeats (LRR) domain (amino acids 674-1300), whose structure apparently bears no resemblance to the RA domains. By a computer algorithm-based search we have unexpectedly found an RA domain in the N-terminal 81 amino acid residues (676-756) of the LRR domain. The purified RA-domain polypeptide exhibits an ability to bind directly to Ras in a GTP-dependent manner and to competitively inhibit Ras-dependent activation of adenylyl cyclase in vitro, with an affinity comparable with that of the whole LRR domain. The specificity of binding of the RA domain to various Ras effector region mutants is indistinguishable from that of the full-length adenylyl cyclase. The activated RAS2 (RAS2(Val-19))-dependent heat shock sensitivity of yeast cells is suppressed by overexpression of the RA domain polypeptide. Further, mutations of the RA domain abolish its Ras binding activity, and adenylyl cyclase molecules carrying these mutations are rendered unactivatable by Ras in vitro. This RA domain bears highest similarity to the Ras-binding domain of Raf-1 based on comparison of its primary and predicted secondary structures with those of other Ras effectors. These results indicate that the RA domain is a primary Ras-binding site for activation of adenylyl cyclase, implicating RA domains as universal modules for interaction of effectors with Ras, conserved from yeast to mammals.

  AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC, Feb. 2002, JOURNAL OF BIOLOGICAL CHEMISTRY, 277(5) (5), 3117 - 3123, English

  [Refereed]

  Scientific journal


• Role of Raf-1 conserved region 2 in regulation of Ras-dependent Raf-1 activation

  H Sendoh, CD Hu, DM Wu, CH Song, Y Yamawaki-Kataoka, J Kotani, T Okada, F Shima, K Kariya, T Kataoka

  Full activation of Raf-l requires the interaction of its CRD with Has. The serine/threonine-rich region, CR2, of Raf-l was implicated in Raf-l regulation, but the underlying mechanism was unclear. Here we show that CRD loses its Ras-binding activity when expressed in connection with CR2, suggesting that CR2 masks CRD. This masking effect is abolished by substitution of Asp or Ala for Ser-259, a growth factor- and TPA-induced phosphorylation site in CR2. Treatment of COS-7 cells expressing Ha-Ras(Val-12) and Raf-l with TPA enhances the Ha-Ras(Val-12)-dependent Raf-l kinase activity. In contrast, the Ha-Ras(Val-12)-dependent activities of the Raf-1(S259D) and Raf-1(S259A) mutants are comparable to that of wild-type Raf-l stimulated by both Ha-Ras(Val-12) and TPA and cannot be further stimulated by TPA treatment. These results suggest that the in vivo phosphorylation of Ser-259 may comprise a crucial step for Ras-dependent Raf-l activation by unmasking CRD and promoting its association with Ras. (C) 2000 Academic Press.

  ACADEMIC PRESS INC, May 2000, BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 271(3) (3), 596 - 602, English

  [Refereed]

  Scientific journal


• Association of yeast adenylyl cyclase with cyclase-associated protein CAP forms a second Ras-binding site which mediates its Ras-dependent activation

  Fumi Shima, Tomoyo Okada, Masahiro Kido, Hiroyoshi Sen, Yasuhiro Tanaka, Masako Tamada, Chang-Deng Hu, Yuriko Yamawaki-Kataoka, Ken-Ichi Kariya, Tohru Kataoka

  Posttranslational modification, in particular farnesylation, of Ras is crucial for activation of Saccharomyces cerevisiae adenylyl cyclase (CYR1). Based on the previous observation that association of CYR1 with cyclase- associated protein (CAP) is essential for its activation by posttranslationally modified Ras, we postulated that the associated CAP might contribute to the formation of a Ras-binding site of CYR1, which mediates CYR1 activation, other than the primary Ras-binding site, the leucine-rich repeat domain. Here, we observed a posttranslational modification-dependent association of Ras with a complex between CAP and CYR1 C-terminal region. When CAP mutants defective in Ras signaling but retaining the CYR1-binding activity were isolated by screening of a pool of randomly mutagenized CAP, CYR1 complexed with two of the obtained three mutants failed to be activated efficiently by modified Ras and exhibited a severely impaired ability to bind Ras, providing a genetic evidence for the importance of the physical association with Ras at the second Ras-binding site. On the other hand, CYR1, complexed with the other CAP mutant, failed to be activated by Ras but exhibited a greatly enhanced binding to Ras. Conversely, a Ras mutant E31K, which exhibits a greatly enhanced binding to the CYR1-CAP complex, failed to activate CYR1 efficiently. Thus, the strength of interaction at the second Ras-binding site appears to be a critical determinant of CYR1 regulation by Ras: too-weak and too-strong interactions are both detrimental to CYR1 activation. These results, taken together with those obtained with mammalian Raf, suggest the importance of the second Ras-binding site in effector regulation.

  American Society for Microbiology, 2000, Molecular and Cellular Biology, 20(1) (1), 26 - 33, English

  Scientific journal


• Characterization of a novel Ras-binding protein Ce-FLI-1 comprising leucine-rich repeats and gelsolin-like domains

  M Goshima, K Kariya, Y Yamawaki-Kataoka, T Okada, M Shibatohge, F Shima, E Fujimoto, T Kataoka

  Ras proteins are conserved from yeasts to mammals and implicated in regulation of the actin cytoskeleton. The flightless-1 (fli-1) gene of Drosophila melanogaster and its homologs in Caenorhabditis elegans and humans encode proteins (FLI-1) comprising a fusion of a leucine-rich repeats (LRRs) domain and a gelsolin-like domain. This LRRs domain is highly homologous to those of three proteins involved in Ras-mediated signaling; Saccharomyces cerevisiae adenylyl cyclase, C. elegans SUR-8, and mammalian RSP-1. Here we report that the LRRs domain of C. elegans FLI-1 (Ce-FLI-1) associates directly with Has (K-d = 11 nM) and, when overexpressed, suppresses the heat shock sensitive phenotype of yeast cells bearing the activated RAS2 gene (RAS2(Val-19)). Further, the gelsolin-like domain of Ce-FLI-1 is shown to possess a Ca2+-independent G-actin-binding activity as well as F-actin-binding and -severing activities. FLI-1 may be involved in regulation of the actin cytoskeleton through Has. (C) 1999 Academic Press.

  ACADEMIC PRESS INC, Apr. 1999, BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 257(1) (1), 111 - 116, English

  [Refereed]

  Scientific journal


• Coiled-coil interaction of N-terminal 36 residues of cyclase-associated protein with adenylyl cyclase is sufficient for its function in Saccharomyces cerevisiae Ras pathway

  Yoshimitsu Nishida, Fumi Shima, Hiroyoshi Sen, Yasuhiro Tanaka, Chie Yanagihara, Yuriko Yamawaki-Kataoka, Ken-Ichi Kariya, Tohru Kataoka

  In the budding yeast Saccharomyces cerevisiae, association with the 70- kDa cyclase-associated protein (CAP) is required for proper response of adenylyl cyclase to Ras proteins. We show here that a small segment comprising the N-terminal 36 amino acid residues of CAP is sufficient for association with adenylyl cyclase as well as for its function in the Ras- adenylyl cyclase pathway as assayed by the ability to confer RAS2(Val-19)- dependent heat shock sensitivity to yeast cells. The CAP-binding site of adenylyl cyclase was mapped to a segment of 119 amino acid residues near its C terminus. Both of these regions contained tandem repetitions of a heptad motif αXXαXXX (where α represents a hydrophobic amino acid and X represents any amino acid), suggesting a coiled-coil interaction. When mutants of CAP defective in associating with adenylyl cyclase were isolated by screening of a pool of randomly mutagenized CAP, they were found to carry substitution mutations in one of the key hydrophobic residues in the heptad repeats. Furthermore, mutations of the key hydrophobic residues in the heptad repeats of adenylyl cyclase also resulted in loss of association with CAP. These results indicate the coiled-coil mechanism as a basis of the CAP- adenylyl cyclase interaction.

  Oct. 1998, Journal of Biological Chemistry, 273(43) (43), 28019 - 28024, English

  [Refereed]

  Scientific journal


• Identification of PLC210, a Caenorhabditis elegans phospholipase C, as a putative effector of Ras

  M Shibatohge, K Kariya, YH Liao, CD Hu, Y Watari, M Goshima, F Shima, T Kataoka

  Mammalian Ras proteins regulate multiple effecters including Raf, Ral guanine nucleotide dissociation stimulator (RalGDS), and phosphoinositide 3-kinase. In the nematode Caenorhabditis elegans, LIN-45 Raf has been identified by genetic analyses as an effector of LET-60 Ras. To search for other effecters in C. elegans, we performed a yeast two-hybrid screening for LET-60-binding proteins. The screening identified two cDNA clones encoding a phosphoinositide-specific phospholipase C (PI-PLC) with a predicted molecular mass of 210 kDa, designated PLC210, PLC210 possesses two additional functional domains unseen in any known PI-PLCs. One is the C-terminal Ras-associating domain bearing a structural homology with those of RalGDS and AF-6. This domain, which could be narrowed down to 100 amino acid residues, associated in vitro with human Ha-Ras in a GTP-dependent manner and competed with yeast adenylyl cyclase for binding Ha-Ras. The binding was abolished by specific mutations within the effector region of Ha-Ras. The other functional domain is the N-terminal CDC25-like domain, which possesses a structural homology to guanine nucleotide exchange proteins for Ras. These results strongly suggest that PLC210 belongs to a novel class of PI-PLC, which is a putative effector of Ras.

  AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC, Mar. 1998, JOURNAL OF BIOLOGICAL CHEMISTRY, 273(11) (11), 6218 - 6222, English

  [Refereed]

  Scientific journal


• Effect of association with adenylyl cyclase-associated protein on the interaction of yeast adenylyl cyclase with Ras protein

  Fumi Shima, Yuriko Yamawaki-Kataoka, Chie Yanagihara, Masako Tamada, Tomoyo Okada, Ken-Ichi Kariya, Tohru Kataoka

  Posttranslational modification of Ras protein has been shown to be critical for interaction with its effector molecules, including Saccharomyces cerevisiae adenylyl cyclase. However, the mechanism of its action was unknown. In this study, we used a reconstituted system with purified adenylyl cyclase and Ras proteins carrying various degrees of the modification to show that the posttranslational modification, especially the farnesylation step, is responsible for 5- to 10-fold increase in Ras-dependent activation of adenylyl cyclase activity even though it has no significant effect on their binding affinity. The stimulatory effect of farnesylation is found to depend on the association of adenylyl cyclase with 70-kDa adenylyl cyclase- associated protein (CAP), which was known to be required for proper in vivo response of adenylyl cyclase to Ras protein, by comparing the levels of Ras- dependent activation of purified adenylyl cyclase with and without bound CAP. The region of CAP required for this effect is mapped to its N-terminal segment of 168 amino acid residues, which coincides with the region required for the in vivo effect. Furthermore, the stimulatory effect is successfully reconstituted by in vitro association of CAP with the purified adenylyl cyclase molecule lacking the bound CAP. These results indicate that the association of adenylyl cyclase with CAP is responsible for the stimulatory effect of posttranslational modification of Ras on its activity and that this may be the mechanism underlying its requirement for the proper in vivo cyclic AMP response.

  American Society for Microbiology, 1997, Molecular and Cellular Biology, 17(3) (3), 1057 - 1064, English

  Scientific journal


• Differential structural requirements for interaction of Ras protein with its distinct downstream effectors

  K Akasaka, M Tamada, F Wang, K Kariya, F Shima, A Kikuchi, M Yamamoto, M Shirouzu, S Yokoyama, T Kataoka

  Ras proteins have multiple effecters of distinct structures that do not share significant structural homology at their Ras interaction sites. To prove possible differences in their recognition mechanisms of Ras, we screened 44 human Ha-Ras proteins carrying mutations in the effector region and its flanking sequences for interaction with human Raf-1, Schizosaccharomyces pombe Byr2, and Saccharomyces cerevisiae adenylyl cyclase. The Ras binding specificities were largely shared between Raf-1 and Byr2 although Ras mutants, Y32F, T35S, and A59E, had their affinities for Byr2 selectively reduced. The only exception was Ras(D38N), which lost the ability to bind Raf-1 while retaining the activity to bind Byr2 and complement the Byr2(-) phenotype of S. pombe. On the other hand, adenylyl cyclase had quite distinct requirements for Ras residues; mutations P34G and T58A selectively abolished the ability to bind and activate it without considerably affecting the interaction with Raf-1 and Byr2. Y32F mutant, whereas losing the ability to activate Raf-1 and Byr2, could activate adenylyl cyclase efficiently. In addition, V45E mutation was found to impair the ability of Ras to activate both Raf-1 and adenylyl cyclase without significantly affecting the binding affinities for them. These results demonstrate that significant differences exist in the recognition mechanisms by which the three effector molecules associate with Ras and suggest that a region of Ras required for activation of the effecters in general may exist separately from that for binding the effectors.

  AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC, Mar. 1996, JOURNAL OF BIOLOGICAL CHEMISTRY, 271(10) (10), 5353 - 5360, English

  [Refereed]

  Scientific journal

• Novel Insights into the Structural Perturbation Induced by the Oncogenic Mutations, Q61L and Q61H, in Ras State 1

  Shigeyuki Matsumoto, Haruka Taniguchi-Tamura, Mitsugu Araki, Takashi Kawamura, Ryo Miyamoto, Chiemi Tsuda, Yasushi Okuno, Fumi Shima, Takashi Kumasaka, Tohru Kataoka

  CELL PRESS, Feb. 2020, BIOPHYSICAL JOURNAL, 118(3) (3), 42A - 42A, English

  Summary international conference


• SACLA,SPring-8並びにNMRを用いた低分子量Gタンパク質RasのGTP加水分解過程における動的構造解析

  佐伯茉帆, 槇野義輝, 松本篤幸, 河村高志, 南後恵理子, 熊坂崇, 島扶美

  2020, 日本分子生物学会年会プログラム・要旨集(Web), 43rd


• Current status of the development of Ras inhibitors

  Fumi Shima, Shigeyuki Matsumoto, Yoko Yoshikawa, Takashi Kawamura, Masayuki Isa, Tohru Kataokat

  Despite the importance of ras as driver genes in many cancers, clinically effective anti-cancer drugs targeting their products, Ras, have been unavailable so far, which was in part ascribable to the apparently 'undruggable' nature of their tertiary structures. Nonetheless, recent studies in academia and industry have identified novel surface pockets accepting small-molecule ligands in both their active GTP-bound and inactive GDP-bound forms (Ras center dot GTP and Ras center dot GDP, respectively), which has led to a surge of investigations into the discovery of Ras-specific inhibitors particularly by utilizing their structural information for structure-based drug design (SBDD). We have been developing Ras inhibitors by SBDD targeting a novel conformation of Ras center dot GTP called state 1, possessing 'druggable' surface pockets, which emerges from the conformational dynamics. In this article, we will survey Ras functions from the structural biological point of view and summarize the current status of the development of Ras inhibitors including our own.

  OXFORD UNIV PRESS, Aug. 2015, JOURNAL OF BIOCHEMISTRY, 158(2) (2), 91 - 99, English

  [Refereed]

  Book review


• rasがん遺伝子産物の新規立体構造情報を利用した分子標的がん治療薬の開発

  Shima Fumi, 熊坂崇, Matsumoto Shigeyuki, Yoshikawa Yoko, 山本雅貴, Kataoka Tohru

  日本放射光学会, 2014, 日本放射光学会誌 放射光, 27(1) (1), 3 - 9, Japanese

  [Refereed]

  Introduction scientific journal


• 【タンパク質修飾による機能変化】 エフェクター活性化過程に必須であるRasの翻訳後修飾 Ras-エフェクター相互作用の高次構造解析

  SHIMA, Fumi, KATAOKA, Toru

  Jun. 2005, 生化学, 77巻, 6号, pp.519-526, Japanese

  Introduction scientific journal


• シクラーゼ結合蛋白質CAPは, Ras蛋白質の翻訳後修飾の出芽酵母アデニル酸シクラーゼ活性化促進効果に必須である.

  島 扶美, 片岡 有里子, 柳原 千枝, 玉田 昌子, 岡田 知代, 苅谷 研一, 片岡 徹

  01 Aug. 1996, 日本分子生物学会年会プログラム・講演要旨集, 19, 314 - 314, Japanese


• Differential structural requirements for interaction of Ras protein with its distinct downstream effectors.

  AKASAKA K, TAMADA M, WANG F, KARIYA K, SHIMA F, KIKUCHI A, YAMAMOTO M, YOKOYAMA S, KATAOKA T

  1996, J. Biol. Chem., 271(10) (10), 5353 - 5360

• ras がん遺伝子産物 Ras を分子標的としたがん治療薬開発の現状

  SHIMA FUMI, Matsumoto Shigeyuki, Yoshikawa Yoko, Kataoka Tohru

  Joint work, がん分子標的治療, 2015, Japanese

  General book


• 生化学 / エフェクター活性化過程に必須であるRasの翻訳後修飾

  SHIMA FUMI

  Joint work, 日本生化学会, Jun. 2005, Japanese

  Scholarly book

• Recognition mechanism of conformational states in RAS by GTPase-acting protein,GAP

  WakakoSakisaka, YoshiteruMakino TaisukeHorikawa, YokoYoshikawa, TakashiKawamura ErikoNngo TakashiKumasaka FumiShima

  第46回日本分子生物学会年会, Dec. 2023, Japanese

  Poster presentation


• Inactivation mechanism og RASelucidated by photoreactive GTP

  HorikawaTaisuke, YoshiteruMakino WakakoSakisaka, YokoYoshikawa, TakashiKawamura ErikoNango, TakashiKumasaka FumiShima

  The 96th Annual Meeting of the Japanese Biochemical Society, Oct. 2023, Japanese

  Oral presentation


• RAS-signaling inhibitors that allosterically disrupt effector conformation and inhibit growth of RAS-driven cancers

  YokoYoshikawa, YoshiteruMakino HirokazuKubota ShigeyukiaMatsumoto, HitomiYuki, TerukiHonma HirooKoyama FumiShima

  THE 82ND ANNUAL MEETING OF THE JAPANESE CANCER ASSOCIATION, Sep. 2023, Japanese

  Oral presentation


• 低分子量Gタンパク質H-Rasのフリーズトラップ結晶構造解析によるGTPase活性分子メカニズムの追跡

  河村高志, 槇野義輝, 長谷川和也, 吉川陽子, 島扶美, 熊坂崇

  第95回日本生化学会大会, Nov. 2022, Japanese

  Poster presentation


• SACLA/SPring-8/NMRを駆使した低分子量G蛋白質Rasの不活性化機構におけるアロステリック構造変化の解明

  槇野義輝, 河村高志, 吉川陽子, 南後恵理子, 熊坂崇, 島扶美

  第95回日本生化学会大会, Nov. 2022, Japanese

  Poster presentation


• SACLA/SPring-8/NMRを用いたRasのアロステリック構造変化の解明

  槇野義輝, 島扶美

  第5回徳島大学統合的がん創薬研究クラスター・合同オンラインミーティング, Mar. 2022, Japanese


• Confoemational changes neighboring the GTP-binding site of small GTPase Ras upon GTP hydrolysis process.

  Maho Saeki, Yoshiteru Makino, Takashi Kawamura, Eriko Nango, Takashi Kumasaka, Fumi Shima

  第94回日本生化学会大会, Nov. 2021

  Poster presentation


• Visualization of enzyme-catalyezed reaction of oncogene product Ras utilizing its photo-reactive substrate

  Fumi Shima, Yoshiteru Makino, Eriko Nango, Takashi Kumasaka

  第94回日本生化学会大会, Nov. 2021, Japanese

  Invited oral presentation


• Analysis of conformational dynamics of small G-proten Ras on GTP hydrolysis process by SACLA,Spring-8 and NMR

  Maho Saeki, Yoshiteru Makino, Shigeyuki Matsumoto, Takashi Kawamura, Eriko Nango, Takashi Kumasaka, Fumi Shima

  第43回日本分子生物学会年会, Dec. 2020, English

  Poster presentation


• -

  Takashi Kawamura , Yoshiteru Makino* , Kazuya Hasegawa , Takanori Nakane , Seiki Baba , Eriko Nango , Rie Tanaka , So Iwata , Takashi Kumasaka and Fumi Shima*

  第33回日本放射光学会年会・放射光科学合同シンポジウム, Jan. 2020


• -

  Fumi Shima* , Yoshiteru Makino

  第3回徳島大学統合的がん創薬研究クラスター合同ミーティング, Dec. 2019

  [Invited]


• Structural changes on GTP hydrolysis of oncogene product Ras revealed by SACLA, Spring-8 and NMR using photo-controllable caged-GTP

  Yoshiteru Makino, Takashi Kawamura, Shigeyuki Matsumoto, Eriko Nango, So Iwata, Takashi Kumasaka, Fumi Shima

  第57回日本生物物理学会年会, Sep. 2019, Japanese, Domestic conference

  Poster presentation


• 構造生物学に基づくがん治療薬の創出研究

  島扶美

  第2回徳島大学統合的がん創薬研究クラスター合同ミーティング, Feb. 2019, 徳島大学統合的がん創薬研究クラスター, 淡路夢舞台 国際会議場

  [Invited]


• Conformational dynamics of small GTPase Ras on GTP hydrolysis process revealed by SACLA, Spring-8 and NMR

  Chika Hagihara, Yoshiteru Makino, Shigeyuki Matsumoto, Takashi Kawamura, Ichiro Mori, Eriko Nango, Takashi Kumasaka, Fumi Shima

  第41回日本分子生物学会年会, Nov. 2018, 日本分子生物学会年会, パシフィコ横浜

  Poster presentation


• Conformational changes on GTP hydrolysis of oncogene product Ras revealed by monitoring of time-dependent NMR signal using caged-GTP

  Chika Hagihara, Yoshiteru Makino, Shigeyuki Matsumoto, Takashi Kawamura, Ichiro Mori, Eriko Nango, Takashi Kumasaka, Tohru Kataoka, Fumi Shima

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


• Novel effect of Ras inhibitor and its application to cancer therapy

  森元 智行, Yoshikawa Yoko, 白川 慶徳, 森 一郎, Kataoka Tohru, Shima Fumi

  2017年度生命科学系学会合同年次大会(第40回日本分子生物学会年会 第90回日本生化学会大会）, Dec. 2017, Japanese, 日本分子生物学会日本生化学会, 神戸, Domestic conference

  Poster presentation


• Ras inhibitors display anti-metastatic effect toward ras-transformed cancer cells via downregulation of lysyl oxidase

  Yoshikawa Yoko, Shima Fumi, Kataoka Tohru

  第76回 日本癌学会学術総会, Sep. 2017, Japanese, 日本癌学会, 横浜, International conference

  Oral presentation


• In silico discovery of small-molecule Ras inhibitors

  Shima Fumi

  University of Washington-Kobe University Joint Symposium on Cell Signaling, Sep. 2015, English, University of Washington and Kobe University, Seattle, USA, International conference

  [Invited]

  Invited oral presentation


• Crystal Mounting Method using Humid Air and Hydrophilic Glue Coating For Ambient and Cryogenic Experiments

  Takashi Kumasaka, Masaki Yamamoto, Kataoka Tohru, Shima Fumi, Matsumoto Shigeyuki, Kawamura Takashi, Nobuo Kamiya, Yasufumi Umena, Naoto Yagi, Seiki Baba

  2015 Annual Meeting of the American Crystallographic Association, Jul. 2015, English, American Crystallographic Association, Philadelphia, USA, International conference

  Oral presentation


• 試料雰囲気湿度調整によるRasタンパク質の結晶内構造転移

  熊坂 崇, 馬場 清喜, 宮野 菜央, Kawamura Takashi, Matsumoto Shigeyuki, 山本 雅貴, Kataoka Tohru, Shima Fumi

  第15回日本蛋白質科学会年会, Jun. 2015, Japanese, 日本蛋白質科学会, 徳島, Domestic conference

  Poster presentation


• GTP結合型H-RasのState 1結晶構造情報に基づく立体構造遷移機構の解明

  Matsumoto Shigeyuki, 宮野 菜央, 馬場 清喜, Liao Jingling, 竹田 あずさ, 山本 雅貴, 熊坂 崇, Kataoka Tohru, Shima Fumi

  第15回日本蛋白質科学会年会, Jun. 2015, Japanese, 日本蛋白質科学会, 徳島, Domestic conference

  Poster presentation


• 創薬支援ネットワークの支援を受けて

  Shima Fumi

  創薬支援ネットワーク・シンポジウム「オールジャパンの創薬支援」創薬立国日本に向けて, Jan. 2015, Japanese, 創薬支援ネットワーク・医薬基盤研究所 創薬支援戦略室, 大阪, Domestic conference

  [Invited]

  Invited oral presentation


• Analysis of the mechanism underlying the anti-metastatic action of Ras inhibitors by using a lung metastatic model mouse

  Osamu Takano, Yoshikawa Yoko, Shima Fumi, Kataoka Tohru

  第72回日本癌学会学術総会, Oct. 2013, English, 日本癌学会, 横浜, Domestic conference

  Poster presentation


• Molecular mechanism of conformational transition of GTP-bound Ras revealed by the crystal structure analysis of M-Ras mutants

  松本 耕祐, Shima Fumi, 村岡 真, 井尻 悠一, 市川 晋也, 荒木 望嗣, 熊坂 崇, 田村 厚夫, Kataoka Tohru

  BMB2010 （第33回日本分子生物学会年回・第83回日本生化学会大会合同大会）, Dec. 2010, Japanese, 日本生化学会大会/日本分子生物学会, 神戸, Domestic conference

  Poster presentation


• GTP結合型M-Ras変異体の二種類の結晶構造から考察される立体構造遷移メカニズム

  松本 耕祐, Shima Fumi, 村岡 真, 井尻 悠一, 市川 晋也, 荒木 望嗣, 熊坂 崇, 田村 厚夫, Kataoka Tohru

  第33回日本分子生物学会年会, Dec. 2010, Japanese, 日本分子生物学会, 神戸, Domestic conference

  Oral presentation


• New strategy for development of anti-cancer drugs targeting the ras oncogene products

  Kataoka Tohru, Shima Fumi

  第69回日本癌学会学術総会, Sep. 2010, Japanese, 日本癌学会, 大阪, Domestic conference

  Invited oral presentation


• ＧＴＰ結合型Ｒａｓの構造遷移における分子メカニズムの解析

  Shima Fumi, Muraoka Shin, Ryo Seirei, Araki Mitsugu, Kataoka Tohru

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

  Poster presentation


• X-ray crystal structure analysis of the budding yeast small GTPase Ras2

  Muraoka Shin, Shima Fumi, Ryo Seirei, Kataoka Tohru

  第82回日本生化学会大会, Oct. 2009, Japanese, 日本生化学会, 神戸, Domestic conference

  Poster presentation


• GTP結合型Ｒａｓの構造遷移メカニズムと創薬への応用

  Shima Fumi

  「Ｇ蛋白質シグナル」研究班会議, Sep. 2009, Japanese, 文部科学省特定領域研究, 千葉県・南房総市, Domestic conference

  Others


• Structural analysis of the novel state1 conformation of Ras protein

  Shima Fumi, Muraoka Shin, Kataoka Tohru

  神戸大学グローバルCOEプログラム「統合的膜生物学の国際教育研究拠点」 第3回ワークショップ, Jul. 2009, Japanese, 神戸大学/グローバルCOE, 淡路, Domestic conference

  Poster presentation


• ＧＴＰ結合型Ｒａｓの立体構造多型性と創薬への応用

  Shima Fumi

  G蛋白質特定領域・膜輸送複合体特定領域合同若手ワークショップ 2009, Jan. 2009, Japanese, 文部科学省特定領域研究, 神戸, Domestic conference

  Oral presentation


• ＧＴＰ結合型M-Ｒａｓの構造遷移におけるH-Ras型アミノ酸置換の影響

  Shima Fumi

  G蛋白質特定領域・膜輸送複合体特定領域合同若手ワークショップ 2009, Jan. 2009, Japanese, 文部科学省特定領域研究, 神戸, Domestic conference

  Oral presentation


• ＧＴＰ結合型H-Ｒａｓの立体構造多型性とエフェクター新規認識機構

  Shima Fumi

  G蛋白質特定領域・膜輸送複合体特定領域合同若手ワークショップ 2009, Jan. 2009, Japanese, 文部科学省特定領域研究, 神戸, Domestic conference

  Oral presentation


• Screening for Ras specific inhibitors based on the conformational equilibrium of Ras in complex with GTP

  Shima Fumi, Muraoka Shin, Sugimoto Takeshi, Yoshikawa Yoko, Kataoka Tohru

  Kobe University Global COE Program International Symposium on Integrative Membrane Biology, Dec. 2008, English, グローバルCOE, 神戸, Domestic conference

  Poster presentation


• GTP結合型H-Rasの高次構造多型性とエフェクターの新規認識機構

  Muraoka Shin, Araki Mitsugu, Yoshikawa Yoko, Ryo Seirei, Kataoka Tohru, Shima Fumi

  特定研究領域「Ｇ蛋白質シグナル」研究班会議, Sep. 2008, Japanese, 特定研究領域「Ｇ蛋白質シグナル」, 新潟, Domestic conference

  Poster presentation


• New strategy to develop Ras specific inhibitors based on conformational equilibrium of Ras oncoprotein.

  Shima Fumi, Muraoka Shin, Sugimoto Takeshi, Yoshikawa Yoko, Kataoka Tohru

  第3回グローバルCOE研究討論会 兼グローバルCOE第2回ワークショップ, Jul. 2008, Japanese, グローバルCOE, 淡路, Domestic conference

  Poster presentation


• New strategy to develop Ras specific inhibitors

  Shima Fumi, Yoshikawa Yoko, Muraoka Shin, Sugimoto Takeshi, Kataoka Tohru

  第3回グローバルCOE研究討論会 兼グローバルCOE第2回ワークショップ, Jul. 2008, Japanese, グローバルCOE, 淡路, Domestic conference

  Poster presentation


• Conformational equilibrium of small GTPases and signal transduction

  Shima Fumi, Ryo Seirei, Muraoka Shin, Yoshikawa Yoko, Kataoka Tohru

  BMB2007第30回日本分子生物学会年会、第80回日本生化学会大会合同大会, Dec. 2007, Japanese, 日本分子生物学会/日本生化学会, 横浜, Domestic conference

  Poster presentation


• New strategy to develop Ras specific inhibitors based on conformational equilibrium of Ras oncoprotein

  Shima Fumi

  2007InternationalSymposiumon“G-proteinSignaling”, Jul. 2007, English, The Japanese Grant-in-Aid for Scientific Research on Priority Areas: “G-protein signal” 文部科学省特定領域研究「Ｇ蛋白質シグナル」, 東京, International conference

  [Invited]

  Invited oral presentation


• Rasをターゲットにした抗癌剤のインシリコ創薬

  Shima Fumi

  第６６回神戸バイオサイエンス研究会, May 2007, Japanese, 神戸バイオサイエンス研究会, 神戸, Domestic conference

  Invited oral presentation


• GTP Dissociation Rate and Conformational Equilibrium in Various Small GTPases

  SHIMA FUMI, MURAOKA SHIN, KATAOKA TOHRU

  日本分子生物学会2006フォーラム「分子生物学の未来」〜コンファレンス&サイエンティフィック・エキジビション〜, Dec. 2006, English, 日本分子生物学会, 名古屋, Domestic conference

  Poster presentation


• Signal transduction and conformational equilibrium in various small GTP-binding proteins

  SHIMA FUMI, KATAOKA TOHRU

  神戸大学21世紀COEプログラム「蛋白質のシグナル伝達機能」 平成18年度第1回研究発表会・若手ポスター発表会, Aug. 2006, Japanese, 神戸大学21世紀COEプログラム「蛋白質のシグナル伝達機能」, 神戸, Domestic conference

  Poster presentation


• Allostery in effector recognition by Ras family small GTPases: conformational transition in the GTP-bound form

  SHIMA FUMI, MURAOKA SHIN, KATAOKA TOHRU

  20th IUBMB International Congress of Biochemistry and Molecular Biology and 11th FAOMBM Congress,第79回日本生化学会大会、第29回日本分子生物学会年会, Jun. 2006, English, The International Union of Biochemistry and Molecular Biology／The The Federation of Asian and Oceanian Biochemists and Molecular Biologists ／日本生化学会／日本分子生物学会, 京都, International conference

  Poster presentation


• Crystal structure of M-Ras reveals a GTP-bound "off" state conformation of Ras family small GTPases

  SHIMA, Fumi, 叶 敏, MURAOKA, Shin, 廖 静伶, 岡本 英嗣, 山本 雅貴, 田村 厚夫, 徳島 大介, 宮川 知也

  Keystone Symposia, Frontiers in Structural Biology, Jan. 2006, English, Keystone Symposia, キーストン, International conference

  Poster presentation


• Crystal structure of M-Ras reveals a GTP-bound "off" state conformation of Ras family small GTPases

  叶 敏, SHIMA, Fumi, MURAOKA, Shin, 廖 静伶, 岡本 英嗣, 山本 雅貴, 田村 厚夫, 徳島 大介, 宮川 知也, KATAOKA, Toru

  第28回日本分子生物学会年会, Dec. 2005, English, 日本分子生物学会, 福岡, Domestic conference

  Poster presentation


• Crystal structure of M-Ras reveals a GTP-bound "off" state conformation of Ras family small GTPase

  廖 静伶, SHIMA, Fumi, MURAOKA, Shin, 叶 敏, 岡本 英嗣, 山本 雅貴, 田村 厚夫, 八木 直人, 徳島 大介, 宮川 知也, KATAOKA, Toru

  第64回日本癌学会学術総会, Sep. 2005, English, 日本癌学会, 札幌, Domestic conference

  Others


• Crystal Structure of the small G protein M-Ras and its implications

  MURAOKA, Shin, 山本 雅貴, 八木 直人, 植木 龍夫, 叶 敏, SHIMA, Fumi, 廖 静伶, 岡本 英嗣, 田村 厚夫, KATAOKA, Toru

  XX Congress of the International Union of Crystallography, Aug. 2005, English, International Union of Crystallography, フィレンツェ, International conference

  Poster presentation


• M-Rasの特異的高次構造とGTPase活性及びｴﾌｪｸﾀｰ認識機

  叶 敏, SHIMA, Fumi, MURAOKA, Shin, 山本 雅貴, 植木 龍夫, 廖 静伶, TAMURA, Takashi, KATAOKA, Toru

  第77回日本生化学会大会, Oct. 2004, English, 日本生化学会, 神戸, Domestic conference

  Poster presentation


• 低分子量GTP結合蛋白質M-RasのX線構造解析

  叶 敏, SHIMA, Fumi, MURAOKA, Shin, 山本 雅貴, 八木 直人, KATAOKA, Toru

  第26回日本分子生物学会年会, Dec. 2003, English, 日本分子生物学会, 横浜, Domestic conference

  Poster presentation

• Development of Novel PPI inhibitors as anti-cancer drugs

  島 扶美

  国立研究開発法人日本医療研究開発機構, AMED創薬支援ネットワーク委託実験調査, Kobe University, Apr. 2024 - Mar. 2025, Principal investigator


• 新規白血病治療薬の開発

  大阪大学医学部付属病院未来医療開発部, 「橋渡し研究推進による」未来医療創出」, 神戸大学, Sep. 2023 - Mar. 2024


• Rasシグナルの網羅的阻害を示す革新的がん治療薬の創出を目指した研究

  国立研究開発法人科学技術振興機構, 研究成果展開事業 社会還元加速プログラムSTART大学・エコシステム推進型 大学推進型, 神戸大学, May 2023 - Mar. 2024, Principal investigator


• Development of Novel PPI inhibitors as anti-cancer drugs

  島 扶美

  国立研究開発法人 日本医療研究開発機構, AMED創薬支援ネットワーク委託実験調査（スクリーニング）, Kobe University, Apr. 2022 - Mar. 2024, Principal investigator


• Elucidation of cancer signal transduction mechanism using photo-controllable Ras on an atomic scale

  島 扶美

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area), Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area), Kobe University, Apr. 2022 - Mar. 2024


• Rasシグナルの網羅的阻害を基軸とした革新的がん治療薬の開発

  島扶美

  国立研究開発法人科学技術振興機構, 産学が連携した研究開発成果の展開 研究成果展開事業 大学発新産業創出プログラム(START) 大学推進型, 神戸大学, Apr. 2022 - Mar. 2023, Principal investigator

  本事業の目的は、神戸大学および大阪工業大学において、5年後までに外部資金の間接経費や寄附金を原資とする継続的なGAPファンドやシード投資ファンドを運営・発展させ、継続的な起業活動支援を可能にすることである。 神戸大学と大阪工業大学に所属する研究者の技術シーズに基づく起業活動支援を通じて技術シーズやビジネスモデルのブラッシュアップを行うとともに、「大学発新産業創出プログラム(START)」の申請やベンチャーキャピタル(VC)へ橋渡しする。同時にさらなる技術シーズの創出につなげることで内閣府事業「スタートアップ・エコシステム拠点都市注)」の「グローバル拠点都市」に採択された「京阪神連携によるスタートアップ・エコシステム拠点形成」に貢献する。 具体的には、神戸大学と大阪工業大学が共同でGAPファンドプログラム、起業活動支援プログラムを構築し、試作品製作、追加データ取得などにより、STARTやVCでの評価や投資判断ができるレベルまでビジネスモデルをブラッシュアップする。


• Development of novel leukemia therapeutics

  島 扶美

  国立研究開発法人日本医療研究開発機構, Translational Research Program, Kobe University, Apr. 2022 - Mar. 2023


• Time-resolved structural analysis of small GTPase Ras signaling transduction by utilizing caged-GTP-coupled Ras protein

  Time-resolved structural analysis of small, GTPase Ras signaling, ransduction by utilizing, caged-GTP-coupled Ras protein

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area), Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area), Kobe University, Oct. 2020 - Mar. 2022

  ① NPEcagedGTP・Ras複合体微結晶に加え、光制御下でのcage離脱速度がより速いpHPcagedGTP を用いたSFXを実施しポケット開閉運動初期のデータ収集を行ために、pHPcagedGTPの合成経路の探索を外注（ナード社）において行った。 cagedATP合成に関する文献と同一、もしくは類似のルート、およびいくつかのルーとを探索したが合成の難易度が比較的高いことが明らかになった。中間体の合成方法は見出されたが、中間体からcagedGTPへの縮合反応が上手く進まない状況にある。現在、グアノシンならびにグアノシンモノリン酸をグアノシン側のパーツとしてして用いた新たな合成ルートを検討している。課題採択ならびに研究開始時期が2020年度下期だったこともあり、pHPcagedGTPの入手には至っていない。 ② NPEcaged-GTP型H-Rasの光照射HSQC_NMR測定とアミノ酸残基ごとのkinetics解析により、GTPの加水分解反応（GDP生成反応）において、２つのSwitch領域の構造変化に先駆けて、当該領域に隣接するα３ヘリックスとP-loopの構造変化が起こることが確認できた。また、GTPの加水分解過程においてRasはState 1構造を経由した後GDP型になることが明らかになった。 ③ GTP加水分解反応に際してState 2構造を経由するかどうかについては、構造変化の速度が溶液中と比較して緩やかになることが予想される、NPEcagedGTP型H-Ras微結晶を用いた光照射・固体31P_ NMR測定が必要と考えらえたため、当該測定を実施したこところ、cage離脱後State 1からState 2を経てGDP型への構造変化を示唆する実験データが得られた。 以上の研究成果を2020.12の第43回日本分子生物学会にて発表した（Saeki et al.）。


• Development of novel Ras-signaling inhibitors

  島 扶美

  国立研究開発法人日本医療研究開発機構, 橋渡し研究戦略的推進プログラム「異分野融合型研究の推進による自立循環型新規医療創出基盤の確立」, 神戸大学, Apr. 2020 - Mar. 2022


• Atomic-scale dynamic structural analysis of Ras by SACLA, which provide promising information on designing cancer therapeutic agents

  Shima Fumi

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C), Grant-in-Aid for Scientific Research (C), Kobe University, Apr. 2019 - Mar. 2022

  Small GTPase Ras functions as a molecular switch by cycling between GTP-bound active and GDP-bound inactive forms in cell signaling pathways. Despite its importance as a cancer driver gene product, dedicated efforts to directly target Ras for decades still have not yielded therapeutic efficacy, due to limited structural information on natural GTP-bound Ras and its ‘undruggable’ nature of drug-binding site, i.e., structural dynamics hampering stable drug-binding, that is intriguingly linked to its GTP hydrolysis activity. Here to elucidate structural dynamics of natural GTP-bound Ras, we performed time-dependent structural analysis of photo-controllable caged-GTP-bound Ras on GTP hydrolysis process by SACLA, SPring-8 and NMR. Photo-irradiation to caged-GTP-bound Ras yielded natural GTP-bound Ras, leading to GTP hydrolysis for production of Ras-GDP. These results provide new insight into the structural dynamics of Ras, realizing novel strategies for development of Ras inhibitors.


• Development of Novel PPI inhibitors as anti-cancer drugs

  島 扶美

  国立研究開発法人 日本医療研究開発機構, AMED創薬支援ネットワーク委託実験調査（標的検証・後期）, 神戸大学, Dec. 2019 - Mar. 2021, Principal investigator

  Competitive research funding


• 抗がん剤の設計基盤となるX線自由電子レーザーによるRasの時分割構造解析

  島 扶美

  公益財団法人ひょうご科学技術協会, 平成31年度学術研究助成, Apr. 2019 - Mar. 2020, Principal investigator

  Competitive research funding


• Ras/Rafシグナル伝達を阻害する新規抗がん剤の研究

  島 扶美

  国立研究開発法人 日本医療研究開発機構, AMED創薬支援ネットワーク委託実験調査, Jul. 2014 - Mar. 2019, Principal investigator

  Competitive research funding


• がん転移治療抗体薬開発のためのインテグリンα10の立体構造解析

  島 扶美

  神戸大学医学部第二内科同門会, 神戸大学医学部第二内科同門会助成金, Apr. 2018 - Dec. 2018, Principal investigator

  Competitive research funding


• 低分子量G蛋白質Rasの翻訳後脂質修飾の役割の構造科学的研究

  片岡 徹

  科学研究費補助金／基盤研究(B), Apr. 2014 - Mar. 2017

  Competitive research funding


• 創薬への応用に向けたrasがん遺伝子産物の構造ダイナミクスの分子機構の解明

  島 扶美

  科学研究費補助金／基盤研究(B), Apr. 2014 - Mar. 2017, Principal investigator

  Competitive research funding


• rasがん遺伝子産物の新規立体構造情報に基づくがん分子標的治療薬の開発

  片岡 徹

  厚生労働科研研究費補助金, Apr. 2011 - Mar. 2016

  Competitive research funding


• 新規Ras機能阻害物質Kobe0065ファミリー化合物のがん転移抑制メカニズムの解析

  島 扶美

  高松宮妃癌研究基金研究助成金, Apr. 2014 - Mar. 2015, Principal investigator

  Competitive research funding


• rasがん遺伝子産物の立体構造情報を基盤としたがん分子標的治療薬の理論設計

  島 扶美

  公益財団法人ひょうご科学技術協会, 学術研究助成金, Apr. 2013 - Mar. 2014, Principal investigator

  Competitive research funding


• Ｒａｓ－Ｒａｆ結合阻害活性を有する抗がん剤の創製

  島 扶美

  学術研究助成基金助成金／基盤研究(C), Apr. 2011 - Mar. 2014, Principal investigator

  Competitive research funding


• ホスホリパーゼＣεの炎症・発癌促進機能に関わる新規シグナル伝達系の解明

  片岡 徹

  科学研究費補助金／基盤研究(B), 2011

  Competitive research funding


• A-STEP「Ras/Rafシグナル伝達を阻害する新規抗がん剤の開発」

  島 扶美

  研究成果最適展開支援プログラム フィージビリティスタディステージ 探索タイプ 拠点型, 2011, Principal investigator

  Competitive research funding


• Ｒａｓの新規高次構造を標的とした抗癌剤の開発

  島 扶美

  科学研究費補助金／基盤研究(C), Apr. 2008 - Mar. 2010, Principal investigator

  Competitive research funding


• 低分子量Ｇ蛋白質の高次構造多型性のシグナル伝達における意義

  島 扶美

  科学研究費補助金／特定領域研究, Apr. 2008 - Mar. 2010, Principal investigator

  Competitive research funding


• 低分子量Ｇ蛋白質の高次構造多型性のシグナル伝達における意義

  島 扶美

  科学研究費補助金／特定領域研究, Apr. 2006 - Mar. 2008, Principal investigator

  Competitive research funding


• 低分子量Ｇ蛋白質のクロストークを仲介するＲａｐ１活性化因子の機能解析

  片岡 徹

  科学研究費補助金／基盤研究(B), 2008

  Competitive research funding


• Ｒａｓ／Ｒａｐ結合（ＲＡ）ドメインによるＧＴＰ結合蛋白質認識機構の解明

  島 扶美

  科学研究費補助金／若手研究(B), Apr. 2004 - Mar. 2006, Principal investigator

  Competitive research funding


• Ｒａｓ／Ｒａｐ標的蛋白質ホスホリパーゼＣεの生体内機能の解明

  片岡 徹

  科学研究費補助金／基盤研究(B), 2005

  Competitive research funding


• 低分子量Ｇ蛋白質による細胞増殖の制御機構

  片岡 徹

  科学研究費補助金／特定領域研究, 2005

  Competitive research funding


• Ｒａｓ／Ｒａｐ結合（ＲＡ）ドメインによるＧＴＰ結合蛋白質認識機構の解明

  島 扶美

  科学研究費補助金／若手研究(B), Apr. 2002 - Mar. 2004, Principal investigator

  Competitive research funding


• Analysis of the Regulatory Mechanism and Function of a Novel Class of Phospholipase C, PLCε

  KATAOKA Tohru, SATOH Takaya, SHIMA Fumi, EDAMATSU Hironori

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research, Grant-in-Aid for Scientific Research (B), Kobe University, 2003 - 2004

  1, We constructed phospholipase C_ε(PLC_ε) knockout mice and showed that the hearts of these mice develop ventricular dilation, which is caused by a volume overload resulting from marked regurgitation and mild stenosis of the semilunar (aortic and pulmonic) valves. We analysed the mechanism of the congenital semilunar valvulogenesis defect causing these phenotypes and concluded that the abnormal thickening and morphology of the semilunar valve leaflets in PLC_ε knockout mice were caused by aberrant cellular proliferation in valve remodeling at the late stages of semilunar valvulogenesis. By analogy with the phenotypes of mice carrying an attenuated epidermal growth factor (EGF) receptor or deficient in heparin-binding EGF, we speculated a crucial role of PLC_ε, as a Ras effector downstream of the EGF receptor, in inhibition of valvular cell proliferation. In fact, we observed increased phosphorylation of Smad1/5/8, which induce valvular cell proliferation downstream of the BMP receptor, in PLC_ε knockout mice. 2, By applying the two stage skin chemical carcinogenesis protocol using DMBA as an initiator and a phorbor ester TPA as a promoter to PLC_ε knockout mice, we showed that PLC_ε plays a crucial role in ras oncogene-induced development of benign tumors as well as in their malignant progression. We further analysed the mechanism of action of PLC_ε. PLC_ε knockout mice showed marked reduction in proliferation of basal layer cells and epidermal hyperplasia induced by TPA, suggesting a crucial role of PLC_ε in downstream signaling from TPA. 3, We disrupted the PLC_ε gene in a model organism Caenorhabditis elegans and observed a sterile phenotype, caused by abnormal contraction of sphincter muscles of the spermatheca. 4, By using BaF3 cells expressing a PDGF receptor mutant lacking the PLC_γ-binding sites, we showed the importance of PH and RA domains by establishing an assay system for Ca^<2+> increase induced by PDGF-dependent PLC_ε activation.


• Analysis of the Function of a Novel Class of Mammalian Phospholipase C, PLCε

  KATAOKA Tohru, EDAMATSU Hironori, SHIMA Fumi, SATOH Takaya

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research, Grant-in-Aid for Scientific Research (B), Kobe University, 2001 - 2002

  We have clarified the regulation mechanism of a novel class of phospholipase C, PLCε, and established PLDε as an effector of small GTPases Ras and Pap1. Also, a physiological function of PlCε has been elucidated. 1. The CDC25 homology domain of PLCε acts as a guanine nucleotide exchange factor for Rap1, thereby amplifying Rap1-dependent signaling. Stimulation of cells by platelet-derived growth factor (PDGF), induces two phase activation of PLCε through activation of Ras and Rap1. The rapid and initial phase of this activation is mediated by Ras at the plasma membrane, whereas Rap1 is responsible for the prolonged activation at the Golgi apparatus. The CDC25 homology domain is crucial for the prolonged activation of PCLε by Rap1. The Ras/Rap1-dependent activation of PLCε prevents BaF3 cells from undergoing apoptosis and sustains their proliferation. 2. Analysis of the spatial and temporal expression patterns of PLCε indicates that in mouse embryos a specific induction of PLCε expression is observed during the course of differentiation of the neural stem cells into the neuronal lineage. In adult, PLCε is expressed abundantly in the heart. The PLCε gene-knockout mice, created by gene targeting, are found to exhibit a phenotype characterized by a market cardiomegaly as well as by overexpression of the heart failure markers as early as 4 weeks of age. Thus, PLCε may play a crucial function in intracellular signaling of the cardiomyocytes by linking the Ras pathway with the Ca^<2+>-calcineurin-NFAT pathway. 3. We have isolated a Caenorhabditis elegans mutant worm lacking the PLCε gene. It exhibits a sterile phenotype due to a disorder in transporting the eggs to the uterus, which is presumably caused by defective relaxation of the sphincters of the spermatheca. It is interesting that both the mammals and the nematodes exhibit a similar phenotype carrying a defect in rhythmic muscular contraction.

• RasRaf binding inhibitors

  Fumi Shima, Yoko, Yoshikawa, Yoshiteru Makino, Hirokazu kubota, Hitomi Yuki, Takashi Kumasaka, Takashi Kawamura

  特願2021-125013, 30 Jul. 2021, Kobe University

  Patent right


• Ras機能阻害作用を有するチオキソチアゾリジン誘導体

  KATAOKA TOHRU, SHIMA FUMI

  特願2013-514037, 09 May 2012, 大学長, 特許6014816, 07 Oct. 2016

  Patent right


• Ras機能阻害作用を有するチオキソチアゾリジン誘導体（アメリカ）

  KATAOKA TOHRU, SHIMA FUMI

  14/116152, 09 May 2012, 大学長, US9056862, 16 Jun. 2015

  Patent right


• THIOXOTHIAZOLIDINE DERIVATIVE HAVING RAS FUNCTION INHIBITORY EFFECT

  SHIMA Fumi

  特願WO 2012/153775 A1, 10 May 2011, 特許WO 2012/153775 A1

  Patent right


• AQUEOUS SOLUTION CONTAING PARTIAL RaS POLYPEPTIDE AND METHOD FOR SCREENING INHIBITOR OF RAS FUNCTION

  SHIMA Fumi

  特願WO/2012/108297 A1, 07 Feb. 2011, 特許WO/2012/108297 A1

  Patent right


• Aqueous Solution Containing Partial Ras Polypeptid and Method for Screening Inhibitor of Ras Function

  SHIMA Fumi

  特願WO2012108297A1, 07 Feb. 2011, 特許WO2012108297A1

  Patent right