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2001 Fiscal Year Final Research Report Summary

Identification of the regulatory elements of human von Willebrand factor for binding to platelet GPlb.

Research Project

Project/Area Number 12670983
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Hematology
Research InstitutionNagoya University

Principal Investigator

MATSUSHITA Tadashi  University Hosoita, Nagoya University, Research Associate, 医学部・附属病院, 助手 (30314008)

Co-Investigator(Kenkyū-buntansha) KOJIMA Tetsuhito  School of Medicine, Nagoya University, Professor, 医学部, 教授 (40161913)
Project Period (FY) 2000 – 2001
Keywordsvon Willebrand factor / GPIh / polymerase chain reaction / platelet / thrombosis / coronary heartdisease / alanine scanning mutagencsis
Research Abstract

In vitro platelet glycoprotein Ib (GPIb) binding of human von Willebrand factor (VWF) increases markedly by exogenous modulators such as ristocetin or botrocetin and the binding does not occur in normal circulation. GPIb binding sites have been assigned in VWF Al domain that consists of a disulfide loop Cysl272 (509l)-Cys1458(695). In contrast, several gain-of-function mutations have been found in two regions comprised of the disulfide loop and its N- and Cterminal flanking regions. In this study, Cysl222(459)-Tyr1271(508), Gin1238(475)Tyrl271 (508), Glu l260(497)-Tyrl27 1(508), and Asp 1459(696)-Asp1472(709) were sequentially deleted of full-length multimeric recombinant VWF. Deletions at either side resulted in normal GPIb binding, indicating that the flanking regions are not GPIb binding sites. However, an additive mutation at Arg1308(545) onto each deletion mutant resulted in the spontaneous GPIb binding without requirements of modulators, suggesting both regions are important for inhibition of GPIb binding. The spontaneous binding was completely inhibited by monoclonal antibodies that recognize the GPIb binding sites. Interestingly, mutations deleted with N-terminal, but not Cterminal flanking regions lost the binding to monoclonal antibodies B328, B710, and 23C7 that selectively inhibit ristocetin-induced GPIb binding and their epitopes were found at His 1268(505) or Asp1269(506). The crystalographic structure of the Al domain suggest that GPIb binding is influenced by the molecular interface between two regions, and the antibody binding to the interface inhibit the binding.

  • Research Products

    (24 results)

All Other

All Publications (24 results)

  • [Publications] T.Sugihara., I.Takahashi., T.Matsushita., et al.: "Identificaion of plasma antibody epitopes and gene abnormalities in Japanese hemophilia A patients with factor VIII inhibitor"Nagoya J Med Sci.. 63. 25-39 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] T.Nakayama., T.Matsushita., et al.: "A case of purpura fuiminans is caused by hornozygous delta8857 mutation (protein C-nagoya) and successfully treated with activated protein C concentrate"Br J Haematol. 110. 727-730 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] T.Matsushita., D.Meyer., J.E.Sadler: "Localization of von Willebrand Factor-binding Sites for Platelet Glycoprotein lb and Botrocetin by Charged-to-Alanine Scanning Mutagenesis"J. Biol. Chem.. 275. 11044-11049 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] A.Katsumi., T.Matsushita., et al.: "Severe factor VII deficiency caused by a novel mutation His348 to Gin in the catalytic domain"Thromb Hacmost. 83. 239-243 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] K.Ishiguro., T.Matsushita. et al.: "Complete antithrombin deficiency in mice results in embryonic lethality"J Clin Invest. 106. 873-878 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Nakayama Y., Matsushita, T., et al.: "Cloning and characterization of the murine antithrombin gene"Thromb Res.. 100. 179-183 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] K.Ishiguro, T.Matsushita, et al.: "Syndecan-4 deficiency leads to high mortality of lipopolysaccharide-injected mice"J Biol Chem.. 276. 47483-47488 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] T.Iwaki, T.Mastushita, et al.: "DNA sequence analysis of protein S deficiency--identification of four point mutations in twelve Japanese subjects"Semin Thromb Hemost. 27. 155-160 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] T.Kojima, T.Matsushita, et al.: "Plasma levels of syndecan-4 (ryudocan) are elevated in patients with acute myocardial infarction"Thromb Haemost. 85. 793-799 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] S.Kunishima, T.Matsushita, et al.: "Mutations in the NMMHC-A gene cause autosomal dominant macrohrombocytopenia with leukocyte inclusions (May-Hegglin anomaly/Sebastian syndrome)"Blood. 97. 1147-1149 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] S.Kunishima, T.Matsushita, et al.: "Identification of six novel MYH9 mutations and genotype-phenotype relationships in autosomal dominant macrothromhocytopenia with leukocyte inclusions"J. Hum. Genet. 46. 722-729 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] M.Yanada, T.Matsushita, et al.: "Impact of antithrombin deficiency in thrombogenesis : lipopolysaccharide and stress-induced thrombus formation in heterozygous antithrombin-deficient mice"Blood. (in press).

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] T. Sugihara. I. Takahashi., T. Matsushita.,etal: "Identification of plasma antibody epitopes and gene abnormalities in Japanese hemophilia A patients with factor VIII inhibitor"Nagoya J Med Sci.. 63. 25-39 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] T. Nakayama., T. Matsushita., etal: "A case of purpura fulminans is aused by homozygous delta8857 mutation (protein C-nagoya) and successfully treated with activated protein C concentrate"Br J Haematol. 110. 727-730 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] T. Matsushita., D. Meyer., J. E. Sadler.: "Localization of von Willebrand Factor-binding Sites for Platelet Glycoprotein Ib and Botrocetin by Charged-to-Alanine Scanning Mutagenesis"J. Biol. Chem. 275. 11044-11049 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] A. Katsumi., T. Matsushita.,etal: "Severe factor VII deficiency caused by a novel mutation His348 to Gin in the catalytic domain."Thromb Haemosi. 83. 239-243 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] K. Ishiguro., T. Matsushita.etal: "Complete antilhrombin deficiency in mice results in embryonic lethality"J Clin Invest. 106. 873-878 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Nakayama Y., Matsushita,T.,etal: "Cloning and characterization of the murine antithrombin gene"Thromb Res. 100. 179-183 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] K. Ishiguro, T., Matsu.shita.clal: "Syndecan-4 deficiency leads to high mortality of lipopolysaccharide-injected mice."J Bioi Chem. 276. 47483-47488 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] T. Iwaki, T., Mastushila,etal: "DNA sequence analysis of' protein S deficiency-identificalion of four point mutations in twelve Japanese subjects."Semin Thromb Hemost. 27. 155-160 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] T. Kojima, T., Matsushita.etal: "Plasma levels of syndecan-4 (ryudocan) are elevated in patients with acute myocardial infarction."Thromb Haemost. 85. 793-799 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] S. Kunishima, T., Matsushita,etal: "Mutations in the NMMHC-A gene cause autosomal dominant macrothrombocytopenia with leukocyte inclusions (May-Hegglin anomaly/Sebastian syndrome)"Blood. 97. 1147-1149 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] S. Kunishima, T., Matsushita.etal: "Identification of six novel MYH9 mutations and genotype-phenotype relationships in autosomal dominant macrothrombocytopenia with leukocyte inclusions"J. Hum. Genet. 46. 722-729 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] M. Yanada, T., Matsushita.etal: "Impact ofantilhrombin deficiency in thrombogenesis : lipopolysaccharide and stress-induced thrombus formation in heterozygous antithrombin-deficient mice"Blood. in press.

    • 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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