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2001 Fiscal Year Final Research Report Summary

Molecular cell pharmacology of novel smooth muscle regulation molecule

Research Project

Project/Area Number 12672212
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field 応用薬理学・医療系薬学
Research InstitutionMie University

Principal Investigator

NAKA Michiko  Mie University, Faculty of Medicine, Assistant, 医学部, 助手 (10093139)

Co-Investigator(Kenkyū-buntansha) TSUNODA Hiroshi  Mie University, Faculty of Medicine, Assistant, 医学部, 助手 (20314114)
NISHIMURA Yuhei  Mie University, Faculty of Medicine, Assistant, 医学部, 助手 (30303720)
Project Period (FY) 2000 – 2001
Keywordssmooth muscle / S100 family / Calcium / Actin / Actin binding gites / calponin / S100C / Actin activated Mg myosin ATPase activity
Research Abstract

S100C is a novel member of the S100 protein family which we purified a novel Ca^<2+>-binding protein and abundant in smooth muscle. We found that S100C inhibits the smooth muscle actin-activated myosin Mg^<2+>-ATPase activity in a dose-dependent manner. Calmodulin and S100L protein did not inhibit actin-activated myosin Mg^<2+>-ATPase activity. S100ao and S100b also did not exhibit any effects. These results suggest that the inhibitory effects of S100C on the actin-activated myosin Mg^<2+>-ATPase activity is specific. Moreover, we investigated whether Ca^<2+>-binding proteins(calmodulin, S100ao, S100b, S100L and S100C) can reverse the inhibitory effects of calponin, a actin-binding protein on the actin-activated myosin Mg^<2+>-ATPase activity. Calmodulin, S100ao and S100b reversed the inhibitory effect of calponin in a Ca^<2+>-dependent manner, S100L had no effect, and S100C had additional inhibitory effects. Furthermore, S100C was found to bind to actin with a stoichiometry of 1 : 6〜7 in the presence of Ca^<2+> and with an affinity of 1 x 10^<-6> M by cosedimentation assays. Moreover, we cross-linked S100C to actin using the carbodiimide zero-length cross-linker 1-ethyl-3-[3-(dimethylamino)propyl] carbodiimide (EDC) in the presence of N-hydroxysuccinimide (NHS). The cross-linked peptides were purified after trypsin digestion, and cross-linked sites were identified by amino acid sequencing. Amino acid sequence analysis indicated that the cross-linking occurred between Lys-191 of actin and Glu-44 of S100C. The results suggest that S100C might be involved in the regulation of actin-activated myosin Mg^<2+>-ATPase activity through its Ca^<2+>-dependent interaction with actin filaments.

  • Research Products

    (12 results)

All Other

All Publications (12 results)

  • [Publications] Xiao Qing Zhao: "Ca2+-dependent Inhibition of Actin-activated Myosin ATPase Activity S100C(S100A11), A Novel Member of the S100 Protein Family"Biochem. Biophys. Res. Commun.. 267. 77-79 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] A.Kondo: "Transcriptional and post-transcriptional regulation of monocyte chemoattractant protein-3 gene expression in human endothelialcellsbyphorbol ester and cAMP signalling"Immunology. 99. 561-568 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] T.Tanaka: "Pharmacogenomics and therapeutic target validation in cerebral vasospasm"J Cardiovasc. Pharm.. 36(Suppl2). S1-S4 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Y.Nishimura: "Calcium-dependent Activation of NFIL3/E4BP4 Gene Expression by Calcineurin/NAT and CaM kinase Signaling"J. Biol. Chem.. 276. 19921-19928 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] H.Suzuki: "The Functional Significance of Heme Oxygenase-1 Gene Induction in a Rat Vasospasm Model"Acta Neurochir [Suppl]. 77. 89-91 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] J.Morikawa: "Molecular Cloning of Novel Mouse and Human Putative Citrate Lyase β Subunit"Biochem. Biophys. Res. Commun.. 289. 1282-1286 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Xiao Qing Zhao, Michiko Naka and Toshio Tanaka: "Ca2+-dependent Inhibition of Actin-activated Myosin ATPase Activity by S100C(100A11), A novel Member of the S100 Protein Family"Biochem. Biophys. Res. Commun.. 267. 77-79 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] A. Kondo, S. Isaji, Y. Nishimura and Toshio Tanaka: "Transcriptional and post-transcriptional regulation of monocyte chemoattractant protein-3 gene expression in human endothelialcellsbyphorbol ester and cAMP signalling"Immunology. 99. 561-568 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] T. Tanaka, Y. Nishimura, H. Tsunoda, M. Naka-: "Pharmacogenomics and therapeutic target validation in cerebral vasospasm"J Cardiovasc. Pharm.. 36(Suppl.2). S1-S4 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Y. Nishimura, T. Tanaka: "Calcium-dependent Activation of NFIL3/E4BP4 Gene Expression by Calcineurin/NFAT and CaM kinase Signaling"J. Biol. chem.. 276(23). 19921-19928 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] H. Suzuki, K. Kanamaru, H. Tsunoda, H. Inada, M. Kuroki, H. Sun, S. Waga, T. Tanaka: "The Functional Significance of Heme Oxygenase-1 Gene Induction in a Rat Vasospasm Model"Acta Neurochir [Suppl]. 77. 89-91 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] J. Morikawa, Y. Nishimura, A. Uchida, T. Tanaka: "Molecular Cloning of Novel Mouse and Human Putative citrate Lyase β-Subunit"Biochem. Biophys. Res. Commun.. 289. 1282-1286 (2001)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 2003-09-17  

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