PURIFICATION AND PHARMACOLOGICAL STUDY OF ENDOTHELIN CONVERTING ENZYME
Project/Area Number |
03670116
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Research Category |
Grant-in-Aid for General Scientific Research (C)
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Allocation Type | Single-year Grants |
Research Field |
General pharmacology
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Research Institution | NATIONAL CARDIOVASCULAR CENTER RESEARCH INSTITUTE |
Principal Investigator |
MATSUSHIMA Yohkazu NATL. CARDIOVASC. CTR. RES. INST. HEAD, 循環動態機能部, 室長 (70165819)
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Project Period (FY) |
1991 – 1992
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Project Status |
Completed (Fiscal Year 1992)
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Budget Amount *help |
¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1992: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1991: ¥1,500,000 (Direct Cost: ¥1,500,000)
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Keywords | ENDOTHELIN / BIG ENDOTHELIN / CONVERTING ENZYME / PHOSPHORAMIDON / EDTA / PEPSTATIN A / EDTA / phosphoramidon / エンドセリン / ビッグエンドセリン / 変換酵素 |
Research Abstract |
1. It is essential for the assessment of endothelin converting enzyme (ECE) activity to measure endothelin-1(ET1). We developed the precise measurement for ET-1 using radioimmunoassay combined with reverse phase HPLC. 2. We examined the effects of ET-1 and big ET-1 on the renal action to study whether or not big ET-1 is converted to ET-1. Intrarenal infusion of ET-1 decreased renal blood flow(RBF), urine output and glomerular filtration rate (GFR), whereas infusion of big ET-1 did not significantly affect RBF, GFR, and urine output, suggesting that big ET-1 could not be converted to mature ET-1. 1992 To characterize the endothelin converting enzyme (ECE) in rat aorta, we prepared the crude membrane fractions obtained by homogenization and centrifugation of homogenate of rat aorta. We observed the ECE activities in the crude membrane fractions which had two optimal pH ; neutral and acidic. Acidic enzyme activity was inhibited by pepstatin A, aspartic protease inhibitor, whereas neutral enzyme was blocked by EDTA, EGTA or phosphoramidon. Further characterization and purification are under investigation.
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Report
(3 results)
Research Products
(3 results)
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[Publications] Akabane S, Yoshimi H, Yoshida K, Kawano Y, Ashida T, Kinoshita O, Matsushima Y, Kawamura M, Imanishi M, Kuramochi M, Omae T: "Differential effects of endothelin-1 and big endothelin on canine kidneys" J. Cardiovasc. Pharmacol. 17(supp1.7). S302-S304 (1991)
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