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Analysis of the Ca^<2+> release mechanism from the E2PCa intermediate of sarcoplasmic reticulum Ca^<2+> pump

Research Project

Project/Area Number 18570119
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Functional biochemistry
Research InstitutionAsahikawa Medical College

Principal Investigator

YAMASAKI Kazuo  Asahikawa Medical College, School of Medicine, Assistant Professor (60241428)

Project Period (FY) 2006 – 2007
Project Status Completed (Fiscal Year 2007)
Budget Amount *help
¥3,890,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥390,000)
Fiscal Year 2007: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2006: ¥2,200,000 (Direct Cost: ¥2,200,000)
KeywordsSarcoplasmic reticulum / Calcium ion / Ca^<2+> pump / Enzyme Kinetics / SERCA1a / SERCA1a
Research Abstract

Development of method for measurement of Ca^<2+> binding to mutants of Ca^<2+> -ATPase, and improvement of its accuracy
Measurement of bound Ca^<2+> was enabled to be done directly by devising the washing solution of the membrane filtoration method in this research, though it was very difficult in the analysis that used the mutant of sarcoplasmic reticulum Ca^<2+>-ATPase to measure Ca^<2+> binding from the low degree of the amount of appearance so far.
Analysis of Ca^<2+> releasing process that uses mutants
In mutant that changes Tyr^<122> of sarcoplasmic reticulum Ca^<2+>-ATPase into Ala (Y122A), I have already found that the E2PCa intermediate accumulates during it in the steady state. In crystal structures of E2P analogs, Tyr^<122> located at the center of the hydrophobic interaction network (Tyr^<122> Hydrophobic cluster: Y122-HC) which situated between cytoplasm domains. The Ala substitution mutants of the other residues include in Y122-HC were made and analyzed its kinetics of Ca^<2 … More +> releasing step. It was revealed from this analysis that the rates of Ca^<2+> release toward lumen side and of association had decreased remarkably compared with a wild type in each mutant. Moreover, the accumulation of the E2PCa in the steady state was observed in a part of mutants as well as Y122A. When Ca^<2+> release from the phosphorylation medium was measured directly by using the above-mentioned method about L119A in this mutant, it was shown that Ca^<2+> release from E2PCa^<2+> was obviously slower than the conversion of the phosphoenzyme (E1PCa_2⇒E2PCa_2). The role of Y122-HC in Ca^<2+> transport of Ca^<2+>-ATPase became clear from the above-mentioned result.
Analysis that uses wild type Ca^<2+>-ATPase
In a series of analysis, it was newly clarified to wild type Ca^<2+>-ATPase that the character to Ca^<2+> of E2P resembled the Y122-HC mutant very much in the absence of K^+. The result that K^+ ion plays an important role to Ca^<2+> release process of Ca^<2+>-ATPase is a finding of an important for recognize the Ca^<2+> transport mechanism of Ca^<2+>-ATPase. Less

Report

(3 results)
  • 2007 Annual Research Report   Final Research Report Summary
  • 2006 Annual Research Report
  • Research Products

    (19 results)

All 2007 2006

All Journal Article (11 results) (of which Peer Reviewed: 4 results) Presentation (8 results)

  • [Journal Article] Critical Role of Glu^<40>-Ser^<48> Loop Linking Actuator Domain and First Transmembrane Helix of Ca^<2+>-ATPase in Ca^<2+> Deocclusion and Release from ADP-insensitive Phosphoenzyme2007

    • Author(s)
      Takashi Daiho
    • Journal Title

      Journal of Biological Chemistry 282

      Pages: 34429-34447

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] Critical Role of ^<40>-Ser^<48> Loop Linking Actuator Domain and First Transmembrane Helix of Ca^<2+>-ATPase in Ca^<2+> Deocclusion and Release from ADP-insensitive Phosphoenzyme2007

    • Author(s)
      Takashi Daiho
    • Journal Title

      Journal of Biological Chemistry vol.282

      Pages: 34429-34447

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Critical Role of Glu^<40>-Ser^<48> Loop Linking Actuator Domain and First Tran smembrane Helix of Ca^<2+>-ATPase in Ca^<2+> Deocclusion and Release from ADP-insensitive Phosphoenzyme2007

    • Author(s)
      Takashi Daiho
    • Journal Title

      Journal of Biological Chemistry 282

      Pages: 34429-34447

    • Related Report
      2007 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Comprehensive Analysis of Expression and Function of 51 Sarco (endo) plasmic Reticulum Ca^<2+>-ATPase Mutants Associated with Darier Disease2006

    • Author(s)
      Yuki Miyauchi
    • Journal Title

      Journal of Biological Chemistry 281

      Pages: 22882-22895

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] ATP2C1 is specifically localized in the basal layer of normal epidermis and its depletion triggers keratinocyte differentiation2006

    • Author(s)
      Masaki Yoshida
    • Journal Title

      Journal of Dermatological Science 43

      Pages: 21-23

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
    • Peer Reviewed
  • [Journal Article] Comprehensive Analysis of Expression and Function of 51 Sarco(endo)plasmic Reticulum Ca^<2+>ATPase Mutants Associated with Darier Disease2006

    • Author(s)
      Yuki Miyauchi
    • Journal Title

      Journal of Biological Chemistry vol.281

      Pages: 22882-22895

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] ATP2Cl is specifically localized in the basal layer of normal epidermis and its depletion triggers keratinocyte differentiation2006

    • Author(s)
      Masaki Yoshida
    • Journal Title

      Journal of Dermatological Science vol.43

      Pages: 21-23

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Journal Article] Comprehensive Analysis of Expression and Function of 51 Sarco(endo)plasmic Reticulum Ca^<2+>-ATPase Mutants Associated with Darier Disease2006

    • Author(s)
      Yuki Miyauchi
    • Journal Title

      Journal of Biological Chemistry 281(32)

      Pages: 22882-22895

    • Related Report
      2006 Annual Research Report
  • [Journal Article] ATP2Cl is specifically localized in the basal layer of normal epidermis and its depletion triggers keratinocyte differentiation2006

    • Author(s)
      Masaki Yoshida
    • Journal Title

      Journal of Dermatological Science 43(1)

      Pages: 21-23

    • Related Report
      2006 Annual Research Report
  • [Journal Article] The Ca^<2+> bound ADP-insensitive Phosphoenzyme of Sarcoplasmic Reticulum Ca^<2+>-ATPase Mutant2006

    • Author(s)
      Kazuo Yamasaki
    • Journal Title

      20^<th> IUBMB Interbational Congress of Biochemistry and Molecular Biology and 11^<th> FAOBMB Congress ABSTRACTS

      Pages: 125-125

    • Related Report
      2006 Annual Research Report
  • [Journal Article] Role of the region linking cytoplasmic and transmembrane domains in processing of phosphorylated intermediates for Ca^<2+> transport of sarcoplasmic reticulum Ca^<2+>-ATPase2006

    • Author(s)
      Takashi Daiho
    • Journal Title

      20^<th> IUBMB Interbational Congress of Biochemistry and Molecular Biology and 11^<th> FAOBMB Congress ABSTRACTS

      Pages: 125-125

    • Related Report
      2006 Annual Research Report
  • [Presentation] 筋小胞体Ca^<2+>-ATPaseのCa^<2+>放出過程におけるTyr122-Hydrophobic clusterとK^+との関連2007

    • Author(s)
      山崎 和生
    • Organizer
      第80回日本生化学会、第30回日本分子生物学会合同大会
    • Place of Presentation
      パシフィコ横浜
    • Year and Date
      2007-12-14
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Annual Research Report 2007 Final Research Report Summary
  • [Presentation] 筋小胞体Ca^<2+>-ATPase変異体によるCa^<2+>を閉塞したADP非感受性リン酸化中間体のアナログの形成2007

    • Author(s)
      大保 貴嗣
    • Organizer
      第80回日本生化学会、第30回日本分子生物学会合同大会
    • Place of Presentation
      パシフィコ横浜
    • Year and Date
      2007-12-14
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Annual Research Report 2007 Final Research Report Summary
  • [Presentation] Relationship between Tyr122-Hydrophobic claster and potassium ion on the Ca^<2+> releasing step of sarcoplasmic reticulum Ca^<2+>-ATPase2007

    • Author(s)
      Kazuo Yamasaki
    • Organizer
      The 30^<th> Annual Meeting of the Molecular Biology Society of Japan and The 80^<th> Annual Meeting of the Japanese Biochemical Society
    • Place of Presentation
      Yokihama
    • Year and Date
      2007-12-14
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Formation of an Analogue for ADP-insensitive Phosphoenzyme with Occluded Ca^<2+> in aMutant of Sarcoplasmic Reticulum Ca^<2+>-ATPase2007

    • Author(s)
      Takashi Daiho
    • Organizer
      The 30^<th> Annual Meeting of the Molecular Biology Society of Japan and The 80^<th> Annual Meeting of the Japanese Biochemical Society
    • Place of Presentation
      Yokohama
    • Year and Date
      2007-12-14
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] The Ca^<2+> bound ADP-insensitive Phosphoenzyme of Sarcoplasmic Reticulum Ca^<2+>-ATPase Mutant2006

    • Author(s)
      Kazuo Yamasaki
    • Organizer
      20th IUBMB Interbational Congress of Biochemistry and Molecular Biology and 11th FAOBMB Congress
    • Place of Presentation
      京都 国際会議場
    • Year and Date
      2006-06-19
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Role of the region linking cytoplasmic and transmembrane domains in processing of phosphorylated intermediates for Ca^<2+> transport of sarcoplasmic reticulum Ca^<2+>-ATPase2006

    • Author(s)
      Takashi Daiho
    • Organizer
      20th IUBMB Interbational Congress of Biochemistry and Molecular Bioloay and 11th FAOBMB Congress
    • Place of Presentation
      京都 国際会議場
    • Year and Date
      2006-06-19
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] The Ca^<2+> bound ADP-insensitive Phosphoenzyme of Sarcoplasmic Reticulum Ca^<2+>-ATPase Mutant2006

    • Author(s)
      Kazuo Yamasaki
    • Organizer
      20^<th> IUBMB International Congress of Biochemistry and Molecular Biology and 11^<th> FAOBMB Congress
    • Place of Presentation
      Kyoto
    • Year and Date
      2006-06-19
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary
  • [Presentation] Role of the region linking cytoplasmic and transmembrane domains in processing of phosphorylated intermediates for Ca^<2+> transport of sarcoplasmic reticulum Ca^<2+>-ATPase2006

    • Author(s)
      Takashi Daiho
    • Organizer
      20^<th> IUBMB International Congress of Biochemistry and Molecular Biology and 11^<th> FAOBMB Congress
    • Place of Presentation
      Kyoto
    • Year and Date
      2006-06-19
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2007 Final Research Report Summary

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Published: 2006-04-01   Modified: 2016-04-21  

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