研究自噬相关TNIP1的蛋白质-蛋白质相互作用。

Investigating Protein-Protein Interactions of Autophagy-Involved TNIP1.

作者信息

Samulevich Michael L, Carman Liam E, Aneskievich Brian J

机构信息

Graduate Program in Pharmacology & Toxicology, University of Connecticut, Storrs, CT, USA.

Department of Pharmaceutical Sciences, School of Pharmacy, University of Connecticut, Storrs, CT, USA.

出版信息

Methods Mol Biol. 2025;2879:63-82. doi: 10.1007/7651_2024_525.

DOI:10.1007/7651_2024_525

PMID:38441723

Abstract

Myriad proteins are involved in the process of autophagy, which they participate in via their protein-protein interactions (PPI). Herein we outline a methodology for examining such interactions utilizing the case of intrinsically disordered protein (IDP) TNIP1 and its interaction with linear M1-linked polyubiquitin. This includes methods for recombinant production, purification, immuno-identification, and analysis of an IDP associated with autophagy, its ordered binding partner, and means of quantitatively analyzing their interaction.

摘要

众多蛋白质参与自噬过程，它们通过蛋白质-蛋白质相互作用（PPI）参与其中。在此，我们概述一种利用内在无序蛋白（IDP）TNIP1及其与线性M1连接的多聚泛素相互作用的情况来检测此类相互作用的方法。这包括重组生产、纯化、免疫鉴定以及分析与自噬相关的IDP、其有序结合伴侣，以及定量分析它们相互作用的方法。