基于稳定同位素标记氨基酸细胞培养法（SILAC）的Sirt1反应性赖氨酸乙酰化蛋白质组定量分析

SILAC-based quantification of Sirt1-responsive lysine acetylome.

作者信息

Chen Yue, Colak Gozde, Zhao Yingming

机构信息

Ben May Department for Cancer Research, University of Chicago, Chicago, IL, USA.

出版信息

Methods Mol Biol. 2013;1077:105-20. doi: 10.1007/978-1-62703-637-5_7.

DOI:10.1007/978-1-62703-637-5_7

PMID:24014402

Abstract

Stable Isotope Labeling by Amino acids in Cell culture (SILAC) is one of the in vivo metabolic labeling methods widely used for dynamic analysis of protein modifications. Here, we describe a general approach to applying SILAC, in combination with affinity enrichment of acetyllysine peptides and mass spectrometry, to study the dynamic changes of the Lysine acetylome in response to Sirt1. The method should be applicable to quantify changes to other post translational modifications in diverse cellular systems.

摘要

细胞培养中氨基酸的稳定同位素标记（SILAC）是广泛用于蛋白质修饰动态分析的体内代谢标记方法之一。在此，我们描述了一种将SILAC与乙酰赖氨酸肽的亲和富集和质谱联用的通用方法，以研究赖氨酸乙酰化组对Sirt1的动态变化。该方法应适用于量化不同细胞系统中其他翻译后修饰的变化。