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Studies of Protein Folding with Gene Manipulation and X-ray Solution Scattering -The Case of Staphylococcal Nuclease-

Research Project

Project/Area Number 02680217
Research Category

Grant-in-Aid for General Scientific Research (C)

Allocation TypeSingle-year Grants
Research Field 生物物性学
Research InstitutionOsaka University (1992)
Tohoku University (1990-1991)

Principal Investigator

KATAOKA Mikio  Osaka University, Faculty of Science, Associate Professor, 理学部, 助教授 (30150254)

Project Period (FY) 1990 – 1992
Project Status Completed (Fiscal Year 1992)
Budget Amount *help
¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 1992: ¥300,000 (Direct Cost: ¥300,000)
Fiscal Year 1991: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1990: ¥1,500,000 (Direct Cost: ¥1,500,000)
KeywordsProtein Folding / Gene Manipulation / X-ray Solution Scattering / Staphylococcal Nuclease / Molten Golbule / Non-native Structure / 蛋白質フォ-ルディング / スタフィロコッカス・ヌクレア-ゼ / 変性 / folding / unfolding / 尿素変性 / Staphylococcal nuclease / 鎖状高分子
Research Abstract

The structural information of unfolded state is essential for the better understanding of protein folding. We have revealed that X-ray solution scattering is useful and unique method to derive the compactness and the globularity of a protein molecule, which are estimated from the radius of gyration and the integral intensity of Kratky plot. The compactness and the globularity are the important factors to describe the extent of folding.
Urea denaturation of Staphylococcal nuclease has been studied by X-ray solution scattering. The denaturation curve obtained from the compactness shows a good agreement with those obtained from CD and fluorescence in the case of wild type nuclease, while the denaturation curve of the mutant belonging to the class m- is different between X-ray and CD or fluorescence. This result strongly suggests that the acquisition of the compactness occurs earlier stage than the formation of the secondary structure.
Staphylococcal nuclease fragment which lacks 13 amino acid residues from its C-terminus is unfolded under a physiological condition, while it is folded upon the addition of the inhibitor, pdTp. The fragment is a good model system for the folding study. The mutant fragments which have amino acid substitutions belonging to class m- are more compact than wild type fragment, while the mutant fragments with amino acid substitution of class m+ are much larger than wild type. The m- fragment takes a different inner structure from native protein. The m+ fragment takes a chain-like conformation. These results indicate that the effect of amino acid substitution is appeared in the unfolded state as well as the folded state.
X-ray solution scattering is successfully applied to the studies of various conformational states of the other proteins.

Report

(4 results)
  • 1992 Annual Research Report   Final Research Report Summary
  • 1991 Annual Research Report
  • 1990 Annual Research Report
  • Research Products

    (42 results)

All Other

All Publications (42 results)

  • [Publications] M.NAKAXAKO: "Quarternary structure of pea phytocrome I dimer stupied with small-angle X-ray scattering and rotary-shadowing electron microscopy" Photochemistry and photobiology. 52. 3-12 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Mikio KATAOKA: "Small-angle X-ray scattering studies of calmodulin mutants with deletions in the linker region of the central helix indicate that the linker has predominantly alpha-helical conformation" Biochemistry. 30. 1188-1192 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Mikio KATAOKA: "Properties of the disorder induced in the purple membrane structure by iodination" Science Report of Tohoku University. 11. 239-250 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Mikio KATAOKA: "Small-angle X-ray scattering study of calmodulin bound to two peptides corresponding to parts of the calmodulin-binding domain of the plasma membrane Ca-pump" Biochemistry. 30. 6247-6251 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Masayoshi NAKASAKO: "Crysallographic characterization by X-ray diffraction of the M-eintermsediate from the phlto-cycle of bacteriorhodopsin at room temperature" FEBS Letters. 292. 73-75 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] 片岡 幹雄: "カルモジュリンの溶液構造" 生物物理. 31. 293-298 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] 片岡 幹雄: "蛋白質の溶液中の構造と動力学" 日本物理学会誌. 46. 1040-1042 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] John M.Flanagan: "Truncated Staphylococcal nuclease is compact but disordered" Proceedings of the National Aceademy of Science U.S.A.89. 748-752 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] 片岡 幹雄: "酸素回折計による蛋白質foldingの研究" Photon Factory News. 9(4). 17-19 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Mikio KATAOKA: "Bacteriorhodopsin reconstituted from two individual helices and the complementary five-helix fragment is photoactive" Photochemistry and Photobiology. 56. 895-901 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Yoshihisa HAGIHAR: "Charge repulsion in the comformational stability of melittin" Biochemistry. 31. 11908-11914 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Fumio TOKUNAGA: "Light-induced structural change of bacteriorhodopsin" Photomedicine and Photobiology. 14. 83-88 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] 片岡 幹雄: "X線溶液散液でみたunfold状態の構造" 生物理. 33. 31-35 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Mikio KATAOKA: "Molten globule of cytochrome c studied by the small-angle X-ray scattering" Journal of molecular Biology. 229. 591-596 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Miko KATAOKA: "Synchrotron Radiation and Life Sciences(eds by H.Sthurmann et al.)" Oxford University Press, (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] M. Nakasako, M. Wada, S. Tokutomi, K. Yamamoto, J. Sakai, M. Kataoka, F. Tokunaga & M. Furuya: "Quarternary structure of pea phytochrome I dimer studied with small-angle X-ray scattering and rotary-shadowing electron microscopy" Photochem. Photobiol. 52. 3-12 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] M. Kataoka, J. Head, A. Persechini, R. Kretsinger & D. Engelman: "Small-angle X-ray scattering studies of calmodulin mutants with deletions in the linker region of the central helix indicate that the linker has a predominantly alpha-helical conformation" Biochemistry. 30. 1188-1192 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] M. Kataoka, N. Sato, T. Hamanaka, Y. Amemiya & F. Tokunaga: "Properties of the disorder induced in the purple membrane structure by iodination" Science Rep. Tohoku Univ. 11. 239-250 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] M. Kataoka, J. Head, T. Vorherr, J. Krebs & E. Carafoli: "Small-angle X-ray scattering study of calmodulin bound to two peptides corresponding to parts of the calmodulin-binding domain of the plasma membrane Ca^<2+>-pump" Biochemistry. 30. 6247-6251 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] M. Nakasako, M. Kataoka, Y. Amemiya & F. Tokunaga: "Crystallographic Characterizatio by X-ray diffraction of the M-intermediate from the photocycle of bacteriorhodopsin at room temperature" FEBS Lett. 292. 73-75 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] M. Kataoka: "Solution structure of calmodulin" Seibutsubutsuri. 31. 293-298 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] M. Kataoka: "Solution structure and dynamics of proteins" Bulletin of Japanese Physical Society. 46. 1040-1042 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] J. Flanagan, M. Kataoka, D. Shortle & D. Engelman: "Truncated Staphylococcal nuclease is compact but disordered" Proc. Natl. Acad. Sci. U. S. A. 89. 748-752 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] M. Kataoka: "Studies of protein folding with the Enzyme Diffractometer" Photon Factory News. 9(4). 17-19 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] M. Kataoka, T. Kahn, Y. Tsujiuchi, D. Engelman & F. Tokunaga: "Bacteriorhodopsin reconstituted from two individual helices and the complementary fivehelix fragment is photoactive" Photochem. Photobiol. 56. 895-901 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] F. Tokunaga, M. Kataoka, K. Mihara & Y. Amemiya: "Light-induced structural change of bacteriorhodopsin" Photomed. Photobiol. 14. 83-88 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Y. Hagihara, M. Kataoka, S. Aimoto & Y. Goto: "Charge repulsion in the conformational stability of melittin" Biochemistry. 31. 11908-11914 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] M. Kataoka: "Structural characterization of a protein unfolded state by solution X-ray scattering" Seibutsubutsuri. 33. 31-35 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] M. Kataoka, Y. Hagihara, K. Mihara & Y. Goto: "Molten globule of cytochrome c studied by the small angle X-ray scattering" J. Mol. Biol. 229. 591-596 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] J.M.Flanagan M.Kataoka,D.Shortle D.M.Fngelman: "Truncated Staphylococcal nuclease is compact but disordered" Proceedings of National Academy of Science,U.S.A.89. 748-752 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] M.Kataoka,T.W.Kahn Y.Tsujiuchi,D.M.Engelman,F.Tokunaga: "Bacteriorhodopsin reconstituted from two individual helices and the complementary five-helix fragment is photoactive" Photochemistry and Photobiology. 56. 895-901 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Y.Hagihara,M.Kataoka S.Aimoto,Y.Goto: "Charge repulsion in the conformational stability of melittin" Biochemistry. 31. 11908-11914 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] 片岡 幹雄: "X線溶液散乱でみたunfold状態の構造" 生物物理. 33. 31-35 (1993)

    • Related Report
      1992 Annual Research Report
  • [Publications] M.Kataoka,Y.Hagihara,K.Mihara Y.Goto: "Molten globule of cytochrome c studied by the smallangle X-ray scattering" Journal of Molecular Biology. (1993)

    • Related Report
      1992 Annual Research Report
  • [Publications] Mikio Kataoka: "Smaliーangle Xーray Scattering Study of Calmodulin Bound to Two Peptides Corresponding to parts of the Calmodulinーbinding Domain of the Plasma Membrane Ca pump" Biochemistry. 30. 6247-6251 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Masayoshi Nakasako: "Crystallographic Characterization by Xーray Diffraction of the Mーintermediate from the Photoーcycle of Bacterioーrhodopsin at Room Temperature" FEBS Letters. 292. 73-75 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] 片岡 幹雄: "カルモジュリンの溶液構造" 生物物理. 31. 293-298 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] 片岡 幹雄: "蛋白質の溶液中での構造と動力学" 日本物理学会誌. 46. 1040-1042 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] John M.Flanagan: "Truncated Staphylococcal Nuclease is Compact but Disordered" Proceedings of National Academy of Science,U.S.A.89. 748-752 (1992)

    • Related Report
      1991 Annual Research Report
  • [Publications] 片岡 幹雄: "酵素四折計による蛋白質foldingの研究" Photon Factory News. 9(4). 17-19 (1992)

    • Related Report
      1991 Annual Research Report
  • [Publications] Mikio Kataoka: "properties of the Disorder Induced in the Purple Membrane Structure by Iodination" Science Reports of the Tohoku University ser 8. 11. 239-250 (1991)

    • Related Report
      1990 Annual Research Report
  • [Publications] Mikio Kataoka: "SmallーAngle Xーray Scattering Studies of Calmodulin Mutants with Deletions in the Linker Region of the Central Helix Indicate That the Linker Region Retains a Predominantly αーhelical Conformation" Biochemistry. 30. 1188-1192 (1991)

    • Related Report
      1990 Annual Research Report

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Published: 1990-04-01   Modified: 2016-04-21  

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