| Project/Area Number |
06556010
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 試験 |
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| Research Field |
蚕糸・昆虫利用学
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| Research Institution | Mie University |
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Principal Investigator |
CHINZEI Yasuo Faculty of Medicin, Mie University, Professor, 医学部, 教授 (60024709)
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| Co-Investigator(Kenkyū-buntansha) |
WATANABE Takashi Nippon Shouji Kaisha Ltd, 部長
KOBAYASHI Jun Faculty of Engineering, Mie University, Assistant, 工学部, 助手 (70242930)
ANDOU Katsuhiko Faculty of Medicin, Mie University, Associate Professor, 医学部, 助教授 (90024710)
三浦 健 三重大学, 医学部, 助手 (60219582)
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| Project Period (FY) |
1994 – 1996
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| Project Status |
Completed (Fiscal Year 1996)
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| Budget Amount *help |
¥5,600,000 (Direct Cost: ¥5,600,000)
Fiscal Year 1996: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1995: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1994: ¥3,900,000 (Direct Cost: ¥3,900,000)
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| Keywords | Rhodnius / salivary gland / anticoagulant / Xase inhibitor / cDNA cloning / baculovirus / expression / vasodilator11. References : / 抗凝血因子 / 内因系凝固因子 / 発現 |
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| Research Abstract |
We have analyzed physiologically active compounds from the salivary glands of blood suckinh insect, Rhodnius prolixus. We found two different anti-coagulant activities from the extract of Rhodnius salivary glands by analyzing activated partial thromboplastin time (APTT) and prothrombin time (PT). Our analyzes showed inhibitory activities of both intrinsic pathway, Xase complex and common pathway, thrombin (factor lla) in the coagulant cascade.
We focussed our analyzes on the Xase inhibitor, and tried to purify and characterize. This inhibitor is a hemeprotein with molecular weight of about 20 kDa. We named it Prolixin-S.Prolixin-S has an inhibitory activity of factor X activation, i.e.Xase complex, which include factor IX,VIII,phospholipid and Ca. We are studying on the mechanism of Xase complex blocking with Prolixin-S.
Prolixin-S cDNA was cloned from Rhodnius salivary gland cDNA library with polyclonal antibody to Prolixin-S,and sequenced. Prolixin-S was expressed by baculovirus insect cell expression system and purified chromatographically. Recominant Prolixin-S had anticoagulant activity as almost same level as native Prolixin-S.rProlixin-S was easily reconstituted as heme-protein with hemine and can be bound nitric oxide (NO) reversiblly. NO bound prolixin-S has a vasodilation activity to rabbit vascular smooth muscle. This molecule, Prolixin-S,is a unique protein which has both anti-coagulant activity and vasodilating activity.
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