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MOLECULAR MECHANISM OF ACTIN S-THIOLATION IN GASTRIC MUCOSAL DEFENSE.

Research Project

Project/Area Number 03454230
Research Category

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

Allocation TypeSingle-year Grants
Research Field Gastroenterology
Research InstitutionSCHOOL OF MEDICINE, UNIVERSITY OF TOKUSHIMA. (1992)
Kyoto Prefectural University of Medicine (1991)

Principal Investigator

ROKUTAN Kazuhito  ASSOCIATE PROFESSOR, DEPARTMENT OF NUTRITION, SCHOOL OF MEDICINE, UNIVERSITY OF TOKUSHIMA., 医学部, 助教授 (10230898)

Co-Investigator(Kenkyū-buntansha) KISHI Kyoichi  PROFESSOR, DEPARTMENT OF NUTRITION, SCHOOL OF MEDICINE, UNIVERSITY OF TOKUSHIMA, 医学部, 教授 (80035435)
AOIKE Akira  ASSOCIATE PROFESSOR, DEPARTMENT OF PREVENTIVE MEDICINE, KYOTO PREFECTURAL UNIVER, 医学部, 助教授 (00117871)
Project Period (FY) 1991 – 1992
Project Status Completed (Fiscal Year 1992)
Budget Amount *help
¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 1992: ¥1,900,000 (Direct Cost: ¥1,900,000)
KeywordsGASTRIC MUCOSAL DEFENSE / CULTURED GASTRIC MUCOSAL CELLS / GLUTATHIONE / ACTIN / S-THIOLATION / 胃粘膜防御機構 / Sーthiolation
Research Abstract

Oxidative stress can induce the formation of protein mixed disulfides with low-molecular-weight thiols, a process termed S-thiolation. It has been suggested that this process might modulate cellular metabolic events under oxidative stress. We analyzed S-thiolation of specific soluble proteins in cultured gastric mucosal cells from guinea pigs by gel electrophoresis and autoradiography after radiolabeling of the intracellular glutathione pool with^<35>S. Hydrogen peroxide (0.2 mM) or diamide (0.2 mM) produced rapid and reversible S-thiolation of a distinct group of proteins with molecular masses of 42, 30, 29, 28, and 22 kilodalton (kD). The extent of S-thiolation in the cells was proportional to the loss of intracellular reduced glutathione and the increase in protein-bound glutathione that occurred during incubation with the agents. Hydrogen peroxide caused prominent S-thiolation of the 30 kD protein within 1 min. Diamide initiated prominent S-thiolation of the 42 kD protein, and the modification persisted over the 20 min study period. This protein was identified as actin by immunoblot analysis and actomyosin precipitation. Fluorescent microscopy revealed that diamide caused a disappearance of normal stress fiber and a concomitant increase in actin polymerization in associated with contraction of the cells. These morphological changes were completely reversible, and the time course was approximately the same as that of the S-thiolation, than dethiolation of actin. In contrast, with cells depleted of glutathione by incubation with buthionine-(R,S)-sulfoximine, the cells detached from the culture plates. S-thiolation of actin could help protect the gastric mucosal cells against irreversible organization of microfilaments under oxidative stress.

Report

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

    (32 results)

All Other

All Publications (32 results)

  • [Publications] Kazuhito Rokutan: "Phagocytosis and stimulation of the respiratory burst by phorbol diester initiates S-thiolation of specific proteins in macrophages." Journal of Immunology. 147. 260-264 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Keiya Nakamura: "Induction of heat shock proteins and their implication in protection against ethanol-induced damage in cultured guinea peg gastric mucosal cells." Gastroenterology. 101. 161-166 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Kazuhito Rokutan: "Specific S-thiolation in gastric mucosal cells:possible role of protein sulfhydryls in gastric cytoprotection." Frontiers in mucosal immunology. 2. 577-580 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] 二木 修司 他: "エタノールのモルモット胃単離壁細胞の酸分泌機構に及ぼす影響" 日本消化器病学会誌. 88. 2094-2100 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] 二木 修司 他: "エタノールによる[Ca^<2+>]i上昇に伴うモルモット単離胃壁細胞の酸分泌機構の変化" 日本消化器病学会誌. 89. 1484-1490 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] 六反 一仁: "Oxidative Stvessにおける細胞骨格の修飾" 医学のあゆみ. 157. 2785-2792 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Kazuhito Rokutan: "Oxidative damage and repair" Kelvin J.A.Davies, 889 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] 六反 一仁: "ストレス研究の新しい展開をめざして" 川井啓市, 244 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] K.Rokutan, J.A.Thomas, & R.B.Johnston: "Jr. Phagocytosis and stimulation of the respiratory burst by phorbol diester initiates S-thiolation of specific proteins in macrophages." J. Immunol. 147. 260-264 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] K.Nakamura, K.Rokutan, N.Marui, A.Aoike, & K.Kawai: "Induction of heat shock proteins and their implication in protection against ethanol-induced damage in cultured guinea pig gastric mucosal cells." Gastroenterol. 101. 161-166 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] S.Niki, K.Nakamura, K.Rokutan, T.Saitoh, A.Aoike, & K.Kawai: "Effect of ethanol on acid secretion by isolated parietal cells from guinea pig." Jpn. J. Gastroenterol. 88. 2094-2100 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] S.Niki, K.Rokutan, K.Nakamura, S.Ogihara, H.Kutsumi, T.Saitoh, A.Aoike & K.Kawai: "Calcium-dependent signaling of acid secretion in isolated parietal cells from guinea pigs and its modulation by ethanol." Jpn. J. Gastroenterol. 89. 1484-1490 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] K.Rokutan, K.Nakamura, N.Marui, A.Aoike, K.Kawai: "Specific S-thiolation in gastric mucosal cells: possible role of protein sulfhydryls in gastric cytoprotection." Frontiers in Mucosal Immunol.2. 577-580 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] K.Rokutan, J.A.Thomas, H.Sies et al: "Role of specific protein S-thiolation under oxidative stress." in Oxidative Damage and Repair. 476-480 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] K.Rokutan: "Modulation of cytoskeletal dynamics under oxidative stress." Igakunoayumi. 157. 270-273 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] K.Rokutan & A.Aoike: "Protein S-thiolation under oxidative stress." Ther. Res. 12. 2785-2792 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] K.Rokutan: "Cytoskeletal dynamics under oxidative stress." Ther. Res. 13. 2537-2538 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] K.Rokutan: "Heat shock proteins in gastric mucosal lesions." Mebio. 19. 66-71 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Kazuhito Rokutan: "Phagocytosis and stimulation of respiratory burst by phorbol diester initiates S-thiolation of specific proteins in macrophages." Journal of Immunology. 147. 260-264 (1991)

    • Related Report
      1992 Annual Research Report
  • [Publications] Keiya Nakamura: "Induction of heat shock proteins and their implication in protection against ethanol-induced damage in cultured guinea pig gastric mucosal cells." Gestroenterology. 101. 161-166 (1991)

    • Related Report
      1992 Annual Research Report
  • [Publications] Kazuhito Rokutan: "Specific S-thiolation in gastric mucosal cells:possible role of protein sulfhydryls in gastric cytoprotection." Frontiers in mucosal immunology. 2. 577-580 (1991)

    • Related Report
      1992 Annual Research Report
  • [Publications] 二木 修司 他: "エタノールのモルモット胃単離壁細胞の酸分泌機構に及ぼす影響" 日本消化器病学会誌. 88. 2094-2100 (1991)

    • Related Report
      1992 Annual Research Report
  • [Publications] 二木 修司 他: "エタノールによる〔Ca_<2+>〕i上昇に伴うモルモット単離胃壁細胞の酸分泌機構の変化" 日本消化器病学会誌. 89. 1484-1490 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] 六反 一仁: "Oxidative stvessにおける細胞骨格の修飾" 医学のあゆみ. 157. 2785-2792 (1991)

    • Related Report
      1992 Annual Research Report
  • [Publications] Kazuhito Rokutan: "Oxidative damage and repair" Kelvin J.A.Davies, 889 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] 六反 一仁: "ストレス研究の新しい展開をめざして" 川井啓市, 244 (1991)

    • Related Report
      1992 Annual Research Report
  • [Publications] Kazuhito Rokutan et al.: "Phagocytosis and汝stimulation of the respiratory burst by phorbol diester initiate Sーthiolation of specific proteins in macrophages." Journal of Immunology. 147. 260-264 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Keiya Nakamura et al.: "Induction of heat shock proteins and their implication in protection against ethanolーinduced damage in cultured guinea pig gastric mucosal cells." Gastroenterology. 101. 161-166 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Kazuhito Rokutan et al.: "Specific Sーthiolation in gastric mucosa;possible role of protein sulfhydryls in gastric cytoprotection." Frontiers of mucosal immunology. 2. 577-580 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] James A.Thomas et al.: "Sーthiolation of protein sulfhydryls" Biological reactive intermediates. 4. 95-103 (1990)

    • Related Report
      1991 Annual Research Report
  • [Publications] 二木 修司他: "エタノ-ルのモルモット胃単離壁細胞の酸分泌機構に及ぼす影響" 日本消化器病学会誌. 88. 2094-2100 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] 六反 一仁: "Oxidative stressにおける細胞骨格の修飾" 医学のあゆみ. 157. 270-273 (1991)

    • Related Report
      1991 Annual Research Report

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Published: 1992-04-01   Modified: 2016-04-21  

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