Investigating Protein-Protein Interactions of Autophagy-Involved TNIP1

Michael L Samulevich; Liam E Carman; Brian J Aneskievich

doi:10.1007/7651_2024_525

Investigating Protein-Protein Interactions of Autophagy-Involved TNIP1

Methods Mol Biol. 2024 Mar 6. doi: 10.1007/7651_2024_525. Online ahead of print.

Authors

Michael L Samulevich¹, Liam E Carman¹, Brian J Aneskievich²

Affiliations

¹ Graduate Program in Pharmacology & Toxicology, University of Connecticut, Storrs, CT, USA.
² Department of Pharmaceutical Sciences, School of Pharmacy, University of Connecticut, Storrs, CT, USA. brian.aneskievich@uconn.edu.

PMID: 38441723
DOI: 10.1007/7651_2024_525

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.

Keywords: Bacterial expression; Heat lysis; Intrinsically disordered protein; Microscale thermophoresis; Nickel affinity chromatography; Recombinant protein.