Ying Yujia, Li Huilin
School of Pharmaceutical Sciences, University of Sun Yat-sen University, No.132 Wai Huan Dong Lu, Guangzhou Higher Education Mega Center, Guangzhou 510006, China; School of Pharmacy, China Pharmaceutical University, 639 Longmian Avenue, Nanjing 211198, China.
School of Pharmaceutical Sciences, University of Sun Yat-sen University, No.132 Wai Huan Dong Lu, Guangzhou Higher Education Mega Center, Guangzhou 510006, China; Guangdong Key Laboratory of Chiral Molecule and Drug Discovery, School of Pharmaceutical Sciences, Sun Yat-sen University, Guangzhou, Guangdong 510006, China.
Methods. 2022 Apr;200:42-57. doi: 10.1016/j.ymeth.2020.06.009. Epub 2020 Jun 13.
Deamidation is a nonenzymatic and spontaneous posttranslational modification (PTM) that introduces changes in both structure and charge of proteins, strongly associated with aging proteome instability and degenerative diseases. Deamidation is also a common PTM occurring in biopharmaceutical proteins, representing a major cause of degradation. Therefore, characterization of deamidation alongside its inter-related modifications, isomerization and racemization, is critically important to understand their roles in protein stability and diseases. Mass spectrometry (MS) has become an indispensable tool in site-specific identification of PTMs for proteomics and structural studies. In this review, we focus on the recent advances of MS analysis in protein deamidation. In particular, we provide an update on sample preparation, chromatographic separation, and MS technologies at multi-level scales, for accurate and reliable characterization of protein deamidation in both simple and complex biological samples, yielding important new insight on how deamidation together with isomerization and racemization occurs. These technological progresses will lead to a better understanding of how deamidation contributes to the pathology of aging and other degenerative diseases and the development of biopharmaceutical drugs.
脱酰胺作用是一种非酶促的自发翻译后修饰（PTM），它会引起蛋白质结构和电荷的变化，与衰老过程中蛋白质组的不稳定性以及退行性疾病密切相关。脱酰胺作用也是生物制药蛋白质中常见的一种PTM，是导致蛋白质降解的主要原因之一。因此，对脱酰胺作用及其相关的修饰（异构化和消旋化）进行表征，对于理解它们在蛋白质稳定性和疾病中的作用至关重要。质谱（MS）已成为蛋白质组学和结构研究中用于位点特异性鉴定PTM的不可或缺的工具。在本综述中，我们重点关注MS分析在蛋白质脱酰胺作用方面的最新进展。特别是，我们提供了关于样品制备、色谱分离和多级质谱技术的最新信息，以便在简单和复杂生物样品中准确可靠地表征蛋白质脱酰胺作用，从而对脱酰胺作用与异构化和消旋化如何共同发生产生重要的新见解。这些技术进步将有助于更好地理解脱酰胺作用如何导致衰老和其他退行性疾病的病理过程以及生物制药药物的开发。
