一种用于检测细胞核相关蛋白磷酸激酶的改良纸质结合法。

A modified paper-binding procedure for the assay of nucleus-associated protein phosphokinases.

作者信息

Goueli S A, Slungaard R, Wilson M J, Ahmed K

出版信息

J Pharmacol Methods. 1980 May;3(3):235-42. doi: 10.1016/0160-5402(80)90004-2.

Abstract

Previously existing paper-binding assay procedures gave results with large variations when employed for the measurement of nucleus-associated protein phosphokinase activities. However, a modified method, utilizing the binding of 32P-labeled phosphoprotein substrates to paper and employing washing procedures in 20% trichloroacetic acid at 60 degrees to 70 degrees C, gave highly reproducible results. This modified procedure was satisfactory with either chromatin or a nonhistone protein fraction derived therefrom as a source of enzyme, and dephosphophosvitin, lysine-rich histones, or casein as phosphoprotein substrates.

摘要

以前存在的纸结合测定程序在用于测量细胞核相关蛋白磷酸激酶活性时，结果差异很大。然而，一种改进的方法，利用32P标记的磷蛋白底物与纸的结合，并在60℃至70℃的20%三氯乙酸中进行洗涤程序，得到了高度可重复的结果。这种改进的程序对于来自染色质或由此衍生的非组蛋白部分作为酶源，以及去磷酸化卵黄高磷蛋白、富含赖氨酸的组蛋白或酪蛋白作为磷蛋白底物来说都是令人满意的。