Protocol for a semi-quantitative approach to identify protein S-palmitoylation in cultured cells by acyl biotin exchange assay

被引：2

作者：

Leishman, Stuart ^{[1
]}

Aljadeed, Najd M. ^{[1
]}

Anand, Paras K. ^{[1
]}

机构：

[1] Imperial Coll London, Dept Infect Dis, London W12 0NN, England

来源：

STAR PROTOCOLS | 2024年 / 5卷 / 02期

基金：

英国医学研究理事会;

关键词：

cell biology; immunology; protein biochemistry;

D O I：

10.1016/j.xpro.2024.103054

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Palmitoylation is a post -translational lipid modification in which palmitic acid is conjugated predominantly to cysteine residues of target proteins, allowing them to tether to cell membranes. Here, we describe a protocol to perform a stepwise acyl biotin exchange assay to identify protein S-palmitoylation. We describe steps for initial blocking of free thiols in protein lysates, subsequent replacement of thioester-linked palmitate groups with a biotin tag for affinity enrichment, and identification of palmitoylated proteins by SDS-PAGE. For complete details on the use and execution of this protocol, please refer to Leishman et al.1

引用

页数：11