凝血酶刺激血小板会导致通过质谱分析所揭示的磷脂酰肌醇5-磷酸增加。
Thrombin stimulation of platelets causes an increase in phosphatidylinositol 5-phosphate revealed by mass assay.
作者信息
Morris J B, Hinchliffe K A, Ciruela A, Letcher A J, Irvine R F
机构信息
Department of Pharmacology, University of Cambridge, Tennis Court Road, CB2 1QJ, Cambridge, UK.
出版信息
FEBS Lett. 2000 Jun 9;475(1):57-60. doi: 10.1016/s0014-5793(00)01625-2.
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-(32)PATP and recombinant PIPkin IIalpha, 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.
磷脂酰肌醇5-磷酸（PtdIns5P）是一种新型肌醇脂质，已被证明是II型磷脂酰肌醇磷酸激酶（PIPkins）的主要底物（Rameh等人，1997年，《自然》390卷，192 - 196页）。采用一种设计用来消除酸性溶剂与塑料制品相互作用的方案，通过新霉素层析从细胞提取物中制备磷脂酰肌醇磷酸（PtdInsP）组分，因为发现这种相互作用会抑制该酶。通过与γ-(32)PATP和重组PIPkin IIα一起孵育，并对形成的放射性标记的磷脂酰肌醇4,5-二磷酸（PtdInsP(2)）进行定量，来测定该组分中的PtdIns5P。该检测方法用于血小板，结果显示在凝血酶刺激10分钟期间，PtdIns5P的质量水平增加，这意味着存在一种激动剂刺激的合成机制。
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