体外分析小RNA蛋白辅因子自组装的技术

Techniques to Analyze sRNA Protein Cofactor Self-Assembly In Vitro.

作者信息

Partouche David, Malabirade Antoine, Bizien Thomas, Velez Marisela, Trépout Sylvain, Marco Sergio, Militello Valeria, Sandt Christophe, Wien Frank, Arluison Véronique

机构信息

Synchrotron SOLEIL, L'Orme des Merisiers Saint Aubin, Gif-sur-Yvette, France.

Université Paris Diderot-Paris 7, Sorbonne Paris Cité, Paris, France.

出版信息

Methods Mol Biol. 2018;1737:321-340. doi: 10.1007/978-1-4939-7634-8_18.

DOI:10.1007/978-1-4939-7634-8_18

PMID:29484601

Abstract

Post-transcriptional control of gene expression by small regulatory noncoding RNA (sRNA) needs protein accomplices to occur. Past research mainly focused on the RNA chaperone Hfq as cofactor. Nevertheless, recent studies indicated that other proteins might be involved in sRNA-based regulations. As some of these proteins have been shown to self-assemble, we describe in this chapter protocols to analyze the nano-assemblies formed. Precisely, we focus our analysis on Escherichia coli Hfq as a model, but the protocols presented here can be applied to analyze any polymer of proteins. This chapter thus provides a guideline to develop commonly used approaches to detect prokaryotic protein self-assembly, with a special focus on the detection of amyloidogenic polymers.

摘要

小调节非编码RNA（sRNA）对基因表达的转录后控制需要蛋白质辅助因子才能发生。过去的研究主要集中在RNA伴侣蛋白Hfq作为辅助因子。然而，最近的研究表明，其他蛋白质可能参与基于sRNA的调控。由于其中一些蛋白质已被证明能自我组装，我们在本章中描述了分析所形成的纳米组装体的实验方案。具体而言，我们将分析重点放在大肠杆菌Hfq作为模型，但这里介绍的实验方案可用于分析任何蛋白质聚合物。因此，本章提供了一个开发常用方法来检测原核生物蛋白质自我组装的指南，特别侧重于检测淀粉样聚合物。