来自大肠杆菌核糖体的50-S亚基。活性核糖体蛋白和蛋白复合物的分离。

50-S subunit from Escherichia coli ribosomes. Isolation of active ribosomal proteins and protein complexes.

作者信息

Wystup G, Teraoka H, Schulze H, Hampl H, Nierhaus K H

出版信息

Eur J Biochem. 1979 Oct;100(1):101-13. doi: 10.1111/j.1432-1033.1979.tb02038.x.

DOI:10.1111/j.1432-1033.1979.tb02038.x

Abstract

A method is described for the isolation of highly purified proteins from the 50-S subunit of Escherichia coli ribosomes. All the proteins from the large subunit could be isolated with the exception of L14, L26, L31 and L34. The isolated proteins are functionally active in reconstituted particles. The method consists of successive NH4Cl/EtOH and LiCl washing steps, which split off distinct groups of proteins from the ribosome. The protein groups are further separated by a combination of gel filtration (Sephadex G-100) and ion-exchange chromatography (carboxymethylcellulose) in the presence of 6 M urea, at neutral pH and 4 degrees C. The purity of the proteins was analyzed by two-dimensional gel electrophoresis. In addition, ten protein complexes were isolated and identified.

摘要

本文描述了一种从大肠杆菌核糖体50-S亚基中分离高纯度蛋白质的方法。除L14、L26、L31和L34外，大亚基中的所有蛋白质均可被分离出来。分离出的蛋白质在重组颗粒中具有功能活性。该方法包括连续的NH4Cl/乙醇和LiCl洗涤步骤，这些步骤可从核糖体上分离出不同的蛋白质组。在6 M尿素存在下，于中性pH和4℃条件下，通过凝胶过滤（Sephadex G-100）和离子交换色谱（羧甲基纤维素）相结合的方法进一步分离蛋白质组。通过二维凝胶电泳分析蛋白质的纯度。此外，还分离并鉴定了十种蛋白质复合物。