用于抗生素研究的枯草芽孢杆菌核糖体蛋白的无标记定量分析

Label-Free Quantitation of Ribosomal Proteins from Bacillus subtilis for Antibiotic Research.

作者信息

Schäkermann Sina, Prochnow Pascal, Bandow Julia E

机构信息

Applied Microbiology, Ruhr-Universität Bochum, Universitätsstr. 150, Bochum, 44801, Germany.

出版信息

Methods Mol Biol. 2017;1520:291-306. doi: 10.1007/978-1-4939-6634-9_18.

DOI:10.1007/978-1-4939-6634-9_18

PMID:27873260

Abstract

Current research is focusing on ribosome heterogeneity as a response to changing environmental conditions and stresses, such as antibiotic stress. Altered stoichiometry and composition of ribosomal proteins as well as association of additional protein factors are mechanisms for shaping the protein expression profile or hibernating ribosomes. Here, we present a method for the isolation of ribosomes to analyze antibiotic-induced changes in the composition of ribosomes in Bacillus subtilis or other bacteria. Ribosomes and associated proteins are isolated by ultracentrifugation and proteins are identified and quantified using label-free mass spectrometry.

摘要

当前的研究聚焦于核糖体异质性，将其作为对不断变化的环境条件和压力（如抗生素压力）的一种反应。核糖体蛋白质化学计量和组成的改变以及其他蛋白质因子的结合是塑造蛋白质表达谱或使核糖体休眠的机制。在此，我们提出一种分离核糖体的方法，用于分析抗生素诱导的枯草芽孢杆菌或其他细菌核糖体组成的变化。通过超速离心分离核糖体及相关蛋白质，并使用无标记质谱法对蛋白质进行鉴定和定量。