Direct Analysis of Protein S-Acylation by Mass Spectrometry.
Direct Analysis of Protein S-Acylation by Mass Spectrometry.
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通过质谱法直接分析蛋白质 S-酰化。
DOI：
10.1007/978-1-4939-9532-5_5
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发表时间：
2019
期刊：
影响因子：
--
通讯作者：
Lin,Cheng
中科院分区：
文献类型：
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作者：
Ji,Yuhuan;Lin,Cheng
动态可逆的蛋白质s -酰化，最常见的是as -棕榈酰化，在蛋白质/膜结合和通过棕榈酰化和去棕榈酰化循环调节细胞内信号传导中起重要作用。质谱（MS）对蛋白质s -酰化的直接分析比间接检测方法有几个好处，因为它可以明确地确定酰基修饰的位置和性质，并且不容易出现错误发现。然而，表征酰基蛋白是具有挑战性的，因为在样品制备和串联质谱分析过程中有酰基损失的趋势。在本章中，我们提出了一种保留不稳定酰基修饰的样品制备方案，以及一种LC-MS/MS工作流程，用于高置信度和灵敏度的s -酰化检测。
Dynamic and reversible proteinS-acylation, most commonly occurring asS-palmitoylation, plays an important role in protein/membrane association and the regulation of intracellular signaling via cycles of palmitoylation and depalmitoylation. Direct analysis of proteinS-acylation by mass spectrometry (MS) offers several benefits over indirect detection methods in that it can definitively determine the location and nature of the acyl modification, and is not prone to false discoveries. However, characterization of acyl proteins is challenging because of the tendency of acyl loss during sample preparation and tandem MS analysis. In this chapter, we present a sample preparation protocol that preserves labile acyl modifications and an LC-MS/MS workflow for detection ofS-acylation with high confidence and sensitivity.