通过温和的核糖核酸酶消化从70-S大肠杆菌核糖体中释放某些核糖体蛋白。

Release of certain ribosomal proteins from 70-S Escherichia coli ribosomes by mild ribonuclease digestion.

作者信息

Gast W H, Leberman R

出版信息

Biochim Biophys Acta. 1976 Apr 15;432(1):98-103. doi: 10.1016/0005-2787(76)90045-9.

Abstract

A method for the release of some proteins from Escherichia coli (MRE 600)ribosomes is described, avoiding extraction with denaturing reagents. High-salt-washed, 70-S ribosomes were treated with pancreatic ribonuclease which led to the release of 12 proteins of the larger ribosomal subunit. Separation of released protein was first attempted by gel filtration and by fractionation with (NH4)2SO4. The number and type of the released proteins were identified by sodium dodecyl sulphate-polyacrylamide gels and two-dimensional gel electrophoresis. The method should prove of use in the large scale purification of proteins L1, L7, and L25.

摘要

本文描述了一种从大肠杆菌（MRE 600）核糖体中释放某些蛋白质的方法，避免使用变性试剂进行提取。用高盐洗涤的70-S核糖体用胰核糖核酸酶处理，这导致了较大核糖体亚基的12种蛋白质的释放。首先尝试通过凝胶过滤和用硫酸铵分级分离来分离释放的蛋白质。通过十二烷基硫酸钠-聚丙烯酰胺凝胶和二维凝胶电泳鉴定释放蛋白质的数量和类型。该方法应证明可用于大规模纯化蛋白质L1、L7和L25。