Protocol for Establishing a Protein-Protein Interaction Network Using Tandem Affinity Purification Followed by Mass Spectrometry in Mammalian Cells

Authors

Weixiang Bian
Hua Jiang
Shan Feng
Junjie Chen
Wenqi Wang
Xu Li

Publication Date

9-16-2022

Journal

STAR Protocols

Abstract

Identification of protein interactors is fundamental to understanding their functions. Here, we describe a modified protocol for tandem affinity purification coupled with mass spectrometry (TAP/MS), which includes two-step purification. We detail the S-, 2×FLAG-, and Streptavidin-Binding Peptide (SBP)- tandem tags (SFB-tag) system for protein purification. This protocol can be used to identify protein interactors and establish a high-confidence protein-protein interaction network based on computational models. This is particularly useful for identifying

Keywords

Animals, Chromatography, Affinity, Mammals, Protein Interaction Maps, Proteins, Tandem Affinity Purification, Tandem Mass Spectrometry

DOI

10.1016/j.xpro.2022.101569

PMID

35874475

PMCID

PMC9304681

PubMedCentral® Posted Date

7-19-2022

PubMedCentral® Full Text Version

Post-print

Published Open-Access

yes