Title:
MUTANT RAS POLYPEPTIDE CRYSTAL
Document Type and Number:
WIPO Patent Application WO/2011/007773
Kind Code:
A1
Abstract:
Disclosed are: co-crystals of GTP, or a GTP analog, and a Ras polypeptide that adopts a three-dimensional conformation having a pocket in the Ras molecular surface; a method for preparing said crystals; and a method for screening for Ras function inhibitors on the basis of three-dimensional conformation information obtained from an X-ray crystallographic analysis of said crystals. Upon observing a Ras mutation that adopts a three-dimensional conformation having a pocket in the molecular surface, mutant Ras polypeptides containing this mutation were obtained, and co-crystals of a GTP analog and said mutant Ras polypeptides were prepared. Further, information about the three-dimensional conformation of said co-crystal, including about the pocket region, was obtained by performing an X-ray crystallographic analysis of said co-crystal.
Inventors:
KATAOKA TOHRU (JP)
SHIMA FUMI (JP)
TAMURA ATSUO (JP)
KUMASAKA TAKASHI (JP)
SHIMA FUMI (JP)
TAMURA ATSUO (JP)
KUMASAKA TAKASHI (JP)
Application Number:
PCT/JP2010/061821
Publication Date:
January 20, 2011
Filing Date:
July 13, 2010
Export Citation:
Assignee:
NAT UNIV CORP UNIV KOBE (JP)
JAPAN SYNCHROTRON RADIATION RES INST (JP)
KATAOKA TOHRU (JP)
SHIMA FUMI (JP)
TAMURA ATSUO (JP)
KUMASAKA TAKASHI (JP)
JAPAN SYNCHROTRON RADIATION RES INST (JP)
KATAOKA TOHRU (JP)
SHIMA FUMI (JP)
TAMURA ATSUO (JP)
KUMASAKA TAKASHI (JP)
International Classes:
C07K14/82; C12N15/09; C30B7/04; C30B29/58; G01N33/15; G01N33/50
Foreign References:
| JP2009165717A | 2009-07-30 |
Other References:
YE M. ET AL: "Crystal structure of M-Ras reveals a GTP-bound ''off'' state conformation of Ras family small GTPases", J BIOL CHEM., vol. 280, no. 35, 2005, pages 31267 - 31275, XP008148843
YANUAR A. ET AL: "Crystal structure of human Rad GTPase of the RGK-family", GENES CELLS, vol. 11, no. 8, 2006, pages 961 - 968, XP008148824
SCHEFFZEK K. ET AL: "The Ras-RasGAP complex: structural basis for GTPase activation and its loss in oncogenic Ras mutants", SCIENCE, vol. 277, no. 5324, 1997, pages 333 - 338, XP008148845
SPOERNER M. ET AL: "Dynamic properties of the Ras switch I region and its importance for binding to effectors", PROC NATL ACAD SCI USA, vol. 98, no. 9, 2001, pages 4944 - 4949, XP008148834
FUMI SHIMA ET AL: "Critical role of posttranslational modification of Ras proteins in effector activation -Ras-Effector Sogo Sayo no Koji Kozo Kaiseki-", JOURNAL OF JAPANESE BIOCHEMICAL SOCIETY, vol. 77, no. 6, 2005, pages 519 - 526, XP008148874
SPLINGARD A. ET AL: "Biochemical and structural characterization of the gem GTPase", J. BIOL. CHEM., vol. 282, no. 3, 2007, pages 1905 - 1915, XP008148838
SPOERNER, M. ET AL., PROC NATL ACAD SCI U S A., vol. 98, no. 9, 24 April 2001 (2001-04-24), pages 4944 - 9
YE M. ET AL., J BIOL CHEM., vol. 280, no. 35, 2 September 2005 (2005-09-02), pages 31267 - 75
SPOERNER M; HERRMANN C; VETTER IR; KALBITZER HR; WITTINGHOFER A, PROC NATL ACAD SCI USA., vol. 98, no. 9, 24 April 2001 (2001-04-24), pages 4944 - 9
CARL BRANDON; JOHNTOOZE ET AL.: "Introduction to Protein Structure", 1992, KYOIKUSYA, pages: 276 - 277
D.E. MCREE: "Practical Protein Crystallography", 1993, ACADEMIC PRESS
ONCOGENE, vol. 9, 1994, pages 2153 - 2157
ONCOGENE, vol. 7, 1992, pages 475 - 480
See also references of EP 2455397A4
YANUAR A. ET AL: "Crystal structure of human Rad GTPase of the RGK-family", GENES CELLS, vol. 11, no. 8, 2006, pages 961 - 968, XP008148824
SCHEFFZEK K. ET AL: "The Ras-RasGAP complex: structural basis for GTPase activation and its loss in oncogenic Ras mutants", SCIENCE, vol. 277, no. 5324, 1997, pages 333 - 338, XP008148845
SPOERNER M. ET AL: "Dynamic properties of the Ras switch I region and its importance for binding to effectors", PROC NATL ACAD SCI USA, vol. 98, no. 9, 2001, pages 4944 - 4949, XP008148834
FUMI SHIMA ET AL: "Critical role of posttranslational modification of Ras proteins in effector activation -Ras-Effector Sogo Sayo no Koji Kozo Kaiseki-", JOURNAL OF JAPANESE BIOCHEMICAL SOCIETY, vol. 77, no. 6, 2005, pages 519 - 526, XP008148874
SPLINGARD A. ET AL: "Biochemical and structural characterization of the gem GTPase", J. BIOL. CHEM., vol. 282, no. 3, 2007, pages 1905 - 1915, XP008148838
SPOERNER, M. ET AL., PROC NATL ACAD SCI U S A., vol. 98, no. 9, 24 April 2001 (2001-04-24), pages 4944 - 9
YE M. ET AL., J BIOL CHEM., vol. 280, no. 35, 2 September 2005 (2005-09-02), pages 31267 - 75
SPOERNER M; HERRMANN C; VETTER IR; KALBITZER HR; WITTINGHOFER A, PROC NATL ACAD SCI USA., vol. 98, no. 9, 24 April 2001 (2001-04-24), pages 4944 - 9
CARL BRANDON; JOHNTOOZE ET AL.: "Introduction to Protein Structure", 1992, KYOIKUSYA, pages: 276 - 277
D.E. MCREE: "Practical Protein Crystallography", 1993, ACADEMIC PRESS
ONCOGENE, vol. 9, 1994, pages 2153 - 2157
ONCOGENE, vol. 7, 1992, pages 475 - 480
See also references of EP 2455397A4
Attorney, Agent or Firm:
SHOJI, Takashi et al. (JP)
Takashi Shoji (JP)
Takashi Shoji (JP)
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