Protein deamidation has been proposed to represent a “molecular clock” that progressively disrupts protein structure and function in human degenerative diseases and natural aging. Importantly, this spontaneous process can also modify therapeutic proteins by altering their purity, stability, bioactivity, and antigenicity during drug synthesis and storage. Deamidation occurs non‐enzymatically in vivo, but can also take place spontaneously in vitro, hence artificial deamidation during proteomic sample preparation can hamper efforts to identify and quantify endogenous deamidation of complex proteomes. To overcome this, mass spectrometry (MS) can be used to conduct rigorous site‐specific characterization of protein deamidation due to the high sensitivity, speed, and specificity offered by this technique. This article reviews recent progress in MS analysis of protein deamidation and discusses the strengths and limitations of common “top‐down” and “bottom‐up” approaches. Recent advances in sample preparation methods, chromatographic separation, MS technology, and data processing have for the first time enabled the accurate and reliable characterization of protein modifications in complex biological samples, yielding important new data on how deamidation occurs across the entire proteome of human cells and tissues. These technological advances will lead to a better understanding of how deamidation contributes to the pathology of biological aging and major degenerative diseases. © 2016 Wiley Periodicals, Inc. Mass Spec Rev 36:677–692, 2017

中文翻译：

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蛋白质脱酰胺质谱分析的最新进展

已经提出蛋白质脱酰胺作用代表“分子钟”，其逐渐破坏人类退行性疾病和自然衰老中的蛋白质结构和功能。重要的是，这种自发过程还可以通过在药物合成和储存过程中改变其纯度，稳定性，生物活性和抗原性来修饰治疗性蛋白质。脱酰胺作用在体内非酶促发生，但也可以在体外自发发生，因此蛋白质组学样品制备过程中的人工脱酰胺作用可能会阻碍鉴定和量化复杂蛋白质组的内源性脱酰胺作用的努力。为了克服这个问题，由于该技术具有很高的灵敏度，速度和特异性，因此可以使用质谱（MS）对蛋白质脱酰胺进行严格的位点特异性表征。本文回顾了蛋白质脱酰胺质谱分析的最新进展，并讨论了常见的“自上而下”和“自下而上”方法的优势和局限性。样品制备方法，色谱分离，质谱技术和数据处理方面的最新进展首次使准确，可靠地表征复杂生物样品中的蛋白质修饰成为可能，产生了重要的新数据，说明脱细胞是如何在整个人类细胞和组织蛋白质组中发生的。这些技术进步将使人们更好地了解脱酰胺作用如何导致生物衰老和主要退行性疾病的病理。©2016 Wiley Periodicals，Inc.质谱Rev 36：677–692，2017