Thrombin stimulation of platelets causes an increase in phosphatidylinositol 5-phosphate revealed by mass assay

被引：84

作者：

Morris, JB ^{[1
]}

Hinchliffe, KA ^{[1
]}

Ciruela, A ^{[1
]}

Letcher, AJ ^{[1
]}

Irvine, RF ^{[1
]}

机构：

[1] Univ Cambridge, Dept Pharmacol, Cambridge CB2 1QJ, England

来源：

FEBS LETTERS | 2000年 / 475卷 / 01期

基金：

英国生物技术与生命科学研究理事会; 英国惠康基金;

关键词：

phosphatidylinositol; 5-phosphate; platelet; thrombin; mass assay; type II phosphatidylinositol phosphate kinase; acid inhibition;

D O I：

10.1016/S0014-5793(00)01625-2

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Phosphatidylinositol 5-phosphate (PtdIns5P), a novel inositol lipid, has been shown to be the major substrate for the type II PtdInsP kinases (PIPkins) [Rameh et al. (1997) Nature 390, 192-196]. A PtdInsP fraction was prepared from cell extracts by neomycin chromatography, using a protocol devised to eliminate the interaction of acidic solvents with plasticware, since this was found to inhibit the enzyme. The PtdIns5P in this fraction was measured by incubating with [gamma-P-32]ATP and recombinant PIPkin II alpha, and quantifying the radiolabelled PtdInsP(2) formed. This assay was used on platelets to show that during 10 min stimulation with thrombin, the mass level of PtdIns5P increases, implying the existence of an agonist-stimulated synthetic mechanism. (C) 2000 Federation of European Biochemical Societies. Published by Elsevier Science B.V. All rights reserved.

引用

页码：57 / 60

页数：4

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