用于昼夜节律分析的小鼠组织的磷酸化蛋白质组和蛋白质组样品制备。

Phosphoproteome and Proteome Sample Preparation from Mouse Tissues for Circadian Analysis.

机构信息

Department of Proteomics and Signal Transduction, Max-Planck Institute of Biochemistry, Martinsried, Germany.

Institute of Medical Psychology, Faculty of Medicine, LMU, Munich, Germany.

出版信息

Methods Mol Biol. 2021;2130:185-193. doi: 10.1007/978-1-0716-0381-9_14.

DOI:10.1007/978-1-0716-0381-9_14

PMID:33284445

Abstract

Recent advances in mass spectrometry (MS)-based quantitative proteomics now allow the identification and quantification of deep proteomes and post-translational modifications (PTMs) in relatively short times. Therefore, in the last few years, this technology has proven successful in the circadian field to characterize temporal oscillations of the proteome and more recently PTMs in cellular systems and in tissues. In this chapter, we describe a robust and simple protocol, based on the EasyPhos workflow, to enable preparation of large number of proteomes and phosphoproteomes from mouse tissues for MS-based quantitative analysis. We additionally discuss computational methods to analyze proteome and phosphoproteome time series to determine circadian oscillations.

摘要

基于质谱（MS）的定量蛋白质组学的最新进展现在可以在相对较短的时间内鉴定和定量深度蛋白质组和翻译后修饰（PTM）。因此，在过去几年中，该技术已在生物钟领域证明是成功的，可用于描述细胞系统和组织中蛋白质组和 PTM 的时间波动。在本章中，我们描述了一种基于 EasyPhos 工作流程的强大而简单的方案，可用于从小鼠组织中制备大量蛋白质组和磷酸蛋白质组，以便进行基于 MS 的定量分析。我们还讨论了用于分析蛋白质组和磷酸蛋白质组时间序列以确定生物钟波动的计算方法。