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Analyses of characterization and structure of NO synthase and NO-carrier protein from blood-sucking insects

Research Project

Project/Area Number 10670227
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field 寄生虫学(含医用動物学)
Research InstitutionMie University

Principal Investigator

YUDA Masao  Mie Univ., Fac. of Med., Assistant, 医学部, 助手 (90293779)

Project Period (FY) 1998 – 1999
Project Status Completed (Fiscal Year 1999)
Budget Amount *help
¥3,700,000 (Direct Cost: ¥3,700,000)
Fiscal Year 1999: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1998: ¥2,800,000 (Direct Cost: ¥2,800,000)
KeywordsCoagulation / inhibitor / Rhodnius / factor IX / Prolixin-S / 吸血性昆虫 / 唾液腺 / 生理活性物質
Research Abstract

Rhodnius prolixus is a blood-sucking Hemipteran insect which is known as a vector of American trypanosomiasis. This insect has several physiologically active molecules in its saliva that assist its blood-sucking behavior. We have already reported purification and the cDNA sequence of a salivary anticoagulant, prolixin-S, from this insect and demonstrated that prolixin-S was an inhibitor of the intrinsic Xase. We also reported that prolixin-S bound nitric oxide (NO) in its heme group as a vasodilator and a platelet aggregation inhibitor.
We further studied on its inhibitory mechanisms of the blood coagulation. We found prolixin-S specifically bound to factor IX/IXa in the presence of CaィイD12+ィエD1 ions. Light scattering and surface plasmon resonance (SPR) studies showed that prolixin-S interfered with factor Ix/IXa binding to the phospholipid membrane, indicating that prolixin-S inhibit Xase activity of factor IXa by interference with its Xase complex formation. Furthermore, reconstitution experiments showed that prolixin-S binding to factor IX strongly inhibited factor IXa generation by factor XIa. We also found that prolixin-S inhibited factor IXa generation by factor VIIa/tissue factor complex and factor IXalfa generation by factor Xa. These results suggests that prolixin-S inhibits both intrinsic and extrinsic coagulations by sequential inhibition of all coagulation pathways in which factor IX participates. It was also suggested that prolixin-S may bind to factor IX/IXa by recognizing conformational change of the Gla domain induced by CaィイD12+ィエD1 binding. To gain further understanding, we are now trying to locate the interactive sites using synthesized Gla-residue peptide.

Report

(3 results)
  • 1999 Annual Research Report   Final Research Report Summary
  • 1998 Annual Research Report
  • Research Products

    (18 results)

All Other

All Publications (18 results)

  • [Publications] P. Waidhet et al.: "Purification and characterization of a thrombin inhibitor from the salivary glands of a malarian vector mosquito, Anopheles stephensi"Biochem. Biophy. Acta.. 1381. 227-233 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Jianxin Sun et al.: "Characterization and cDNA cloning of a hemoprotein in the salivary glands of the blood-sucking insect, Rhodinius prolixus"Insect Biochem. Mol. Biol.. 28. 191-200 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Yuji Kaneko et al.: "Kinetic analysis on nitric oxide binding of recombinant Prolixin-S, a nitric oxide transport protein from the bug, Rhodnius prolixus"Biochemica et Biophysica Acta.. 1431. 492-499 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Jianxin Sun et al.: "Changes in salivary gland proteins during blood feeding in Rhodnius prolixus, and dynamics of antibodies specific to salivary antigens"Med. Entomol. Zool.. 50. 41-50 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Yuji Kaneko et al.: "Effects of recombinant nitrophorin-2 nitric oxide comples on vascular smooth muscle"Biosci. Biotech. Biochem.. 63. 1488-1490 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Haruhiko Isawa et al.: "The insect salivary protein, prolixin-S, inhibits factor lXa generation and Xase complex formation in the bood coagulation pathway"J. Biol. Chem.. 275. 6636-6641 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Philomene Waidohet-Kouadio, Masao Yuda, Katsuhiko Ando and Yasuo Chinzei: "Purification and characterization of a thrombin inhibitor from the salivary glands of a malarian vector mosquito, Anopheles stephensi"Biochem. Biophy. Acta.. 1381. 227-233 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Jianxin Sun, Masao Yuda, Ken Miura and Yasuo Chinzei: "Characterization and cDNA cloning of a hemoprotein in the salivary glands of the blood-sucking insect, Rhodnius prolixus"Insect Biochem. Mol. Biol.. 28. 191-200 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Yuji Kaneko, Masao Yuda, Takayoshi Iio, Tomohiko Murase and Yasuo Chinzei: "Kinetic analysis on nitric oxide binding of recombinant Prolixin-S, a nitric oxide transport protein from the blood-sucking bug, Rhodnius prolixus"Biochemica et Biophysica. Acta.. 1431. 492-499 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Jianxin Sun, Philomene Waidohet-Kouadio, Katsuhiko Ando and Yasuo Chinzei: "Changes in salivary gland proteins during blood feeling in Rhodnius prolixus, and dynamics of antibodies specific to salivary antigens in rabbits bitten by the bugs"Med. Entomol. Zool.. 50. 41-50 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Yuji Kaneko, Hideki Shojo, Masao Yoda and Yasuo Chinzei: "Effects of recombinant nitrophorin-2 nitric oxide compels vascular smooth muscle"Biosci. Biotech. Biochem.. 63. 1488-1490 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] Haruhiko Isawa, Masao Yuda, Kentaro Yoneda and Yasuo Chinzei: "The insect salivary protein, Prolixin-S, inhibits factor IXa generation and Xase complex formation in the blood coagulation pathway"J. Biol. Chem.. 275. 6636-6641 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1999 Final Research Report Summary
  • [Publications] P.Waidhet et al.: "Purification and characterization of a thrombin inhibitor from the salivary glands of a malarian vector mosquito,Anopheles stephensi"Biochem.Biophy.Acta.. 1381. 227-233 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Jianxin Sun et al.: "Characterization and cDNA cloning of a hemoprotein in the salivary glands of the blood-sucking insect,Rhodnius prolixus"Insect Biochem.Mol.Biol.. 28. 191-200 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Yuji Kaneko et al.: "Kinetic analysis on nitric oxide binding of recombinant Prolixin-S,a nitric oxide transport protein from the bug,Rhodnius prolixus"Biochemica et Biophysica Acta.. 1431. 492-499 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Jianxin Sun et al.: "Changes in salivary gland proteins during blood feeding in Rhodnius prolixus,and dynamics of antibodies specific to salivary antigens"Med.Entomol.Zool.. 50. 41-50 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Yuji Kaneko et al.: "Effects of recombinant nitrophorin-2 nitric oxide comples on vascular smooth muscle"Biosci.Biotech.Biochem.,. 63. 1488-1490 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Haruhiko Isawa et al.: "The insect salivary protein,prolixin-S,inhibits factor Ixa generation and Xase complex formation in the blood coagulation pathway"J.Biol.Chem.. 275. 6636-6641 (2000)

    • Related Report
      1999 Annual Research Report

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Published: 1998-04-01   Modified: 2016-04-21  

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