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鸡突触后致密物中一些新碱性蛋白的检测与特性分析

Detection and characterization of some new basic proteins in chicken postsynaptic densities.

作者信息

Yang Z W, Babitch J A

出版信息

J Neurochem. 1986 Jul;47(1):282-90. doi: 10.1111/j.1471-4159.1986.tb02860.x.

Abstract

Chicken brain postsynaptic density (PSD) polypeptides, obtained by treating synaptosomes with 0.5% Triton X-100 and then further purified on a sucrose gradient, are demonstrated to contain four basic proteins of 76K (pI greater than 9.2), 58K (pI 8.1-8.8, heterogeneous), 40K (pI 9.0), and 24K (pI 8.9). Nonequilibrium pH gradient-sodium dodecyl sulfate two-dimensional gels further reveal six more basic proteins with pI values higher than 9.2: 76K, 52K, 47K, 45K, 36K, and 34K. These basic proteins are a major part of the total chicken PSD polypeptides appearing on the gels. Some of these basic proteins (58K, 52K, 47K, 36K, 24K, and two at 76K) are distinguishable from those of brain mitochondria, the major contaminant. The 40K and 34K proteins may be common mitochondrial polypeptides. The 45K protein is probably a mitochondrial contaminant. A number of proteins including 76K (synapsin I-like protein) and 58K, along with some other minor ones, can be phosphorylated by endogenous protein kinase(s) in the presence of Ca2+, Mg2+, and [gamma-32P]ATP. No PSD basic proteins bind Ca2+.

摘要

鸡脑突触后致密物(PSD)多肽是通过用0.5% Triton X-100处理突触小体,然后在蔗糖梯度上进一步纯化而获得的,结果表明其含有四种碱性蛋白,分子量分别为76K(pI大于9.2)、58K(pI 8.1 - 8.8,不均一)、40K(pI 9.0)和24K(pI 8.9)。非平衡pH梯度-十二烷基硫酸钠二维凝胶进一步揭示出另外六种pI值高于9.2的碱性蛋白:76K、52K、47K、45K、36K和34K。这些碱性蛋白是凝胶上出现的总鸡PSD多肽的主要部分。其中一些碱性蛋白(58K、52K、47K、36K、24K以及两种76K的蛋白)可与主要污染物脑线粒体的蛋白区分开来。40K和34K蛋白可能是常见的线粒体多肽。45K蛋白可能是线粒体污染物。包括76K(突触素I样蛋白)和58K在内的许多蛋白,以及其他一些次要蛋白,在Ca2 +、Mg2 +和[γ-32P]ATP存在的情况下可被内源性蛋白激酶磷酸化。没有PSD碱性蛋白能结合Ca2 +。

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