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一种基于玻连蛋白被蛋白激酶A选择性磷酸化的酶活性测定方法。

An enzymatic assay for vitronectin based on its selective phosphorylation by protein kinase A.

作者信息

Korc-Grodzicki B, Chain D, Kreizman T, Shaltiel S

机构信息

Department of Chemical Immunology, Weizmann Institute of Science, Rehovot, Israel.

出版信息

Anal Biochem. 1990 Aug 1;188(2):288-94. doi: 10.1016/0003-2697(90)90608-c.

Abstract

The catalytic subunit (C) of cAMP-dependent protein kinase selectively phosphorylates vitronectin, a plasma protein that promotes cell adhesion and platelet aggregation, inhibits the inactivation of thrombin by antithrombin III, and participates in complement function. This specific phosphorylation is used here (a) to develop an enzymatic assay for vitronectin (with C and [gamma-32P]ATP) which can be used to identify the vitronectin-containing fractions at each stage of its purification; (b) to radioactively label vitronectin and differentiate between the intact and the nicked form of this protein in structure-function studies; and (c) to identify possible vitronectin-related proteins in the plasma of other animal species.

摘要

环磷酸腺苷依赖性蛋白激酶的催化亚基(C)可选择性地磷酸化玻连蛋白,玻连蛋白是一种血浆蛋白,可促进细胞黏附和血小板聚集,抑制抗凝血酶III对凝血酶的灭活,并参与补体功能。本文利用这种特异性磷酸化作用:(a)开发一种玻连蛋白的酶促测定法(使用C和[γ-32P]ATP),可用于在其纯化的每个阶段鉴定含玻连蛋白的组分;(b)对玻连蛋白进行放射性标记,并在结构功能研究中区分该蛋白的完整形式和切口形式;以及(c)鉴定其他动物物种血浆中可能与玻连蛋白相关的蛋白质。

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