人外泌体及其辅助因子复合物的亲和蛋白质组学分析

Affinity Proteomic Analysis of the Human Exosome and Its Cofactor Complexes.

作者信息

Winczura Kinga, Domanski Michal, LaCava John

机构信息

School of Biosciences, College of Life and Environmental Sciences, University of Birmingham, Birmingham, UK.

Department of Chemistry and Biochemistry, University of Bern, Bern, Switzerland.

出版信息

Methods Mol Biol. 2020;2062:291-325. doi: 10.1007/978-1-4939-9822-7_15.

DOI:10.1007/978-1-4939-9822-7_15

PMID:31768983

Abstract

In humans, the RNA exosome consists of an enzymatically inactive nine-subunit core, with ribonucleolytic activity contributed by additional components. Several cofactor complexes also interact with the exosome-these enable the recruitment of, and specify the activity upon, diverse substrates. Affinity capture coupled with mass spectrometry has proven to be an effective means to identify the compositions of RNA exosomes and their cofactor complexes: here, we describe a general experimental strategy for proteomic characterization of macromolecular complexes, applied to the exosome and an affiliated adapter protein, ZC3H18.

摘要

在人类中，RNA外切体由一个无酶活性的九亚基核心组成，其核糖核酸酶活性由其他组分提供。几种辅助因子复合物也与外切体相互作用——这些复合物能够招募不同的底物并确定其活性。亲和捕获结合质谱分析已被证明是鉴定RNA外切体及其辅助因子复合物组成的有效方法：在此，我们描述了一种用于大分子复合物蛋白质组学表征的通用实验策略，并将其应用于外切体和一种附属衔接蛋白ZC3H18。