GTP‐binding proteins are among the most important enzyme families that are involved in a plethora of biological processes. However, owing to the enormous diversity of the nucleotide‐binding protein family, comprehensive analyses of the expression level, structure, activity, and regulatory mechanisms of GTP‐binding proteins remain challenging with the use of conventional approaches. The many advances in mass spectrometry (MS) instrumentation and data acquisition methods, together with a variety of enrichment approaches in sample preparation, render MS a powerful tool for the comprehensive characterizations of the activities and expression levels of various GTP‐binding proteins. We review herein the recent developments in the application of MS‐based techniques, together with general and widely used affinity enrichment approaches, for the proteome‐wide and targeted capture, identification, and quantification of GTP‐binding proteins. The working principles, advantages, and limitations of various strategies for profiling the expression level, activity, posttranslational modifications, and interactome of GTP‐binding proteins are discussed. It can be envisaged that future applications of MS‐based proteomics will lead to a better understanding about the roles of GTP‐binding proteins in different biological processes and human diseases. © 2020 John Wiley & Sons Ltd. Mass Spec Rev.

中文翻译：

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通过质谱法对生物样品中的 GTP 结合蛋白进行全球和有针对性的分析

GTP 结合蛋白是参与众多生物过程的最重要的酶家族之一。然而，由于核苷酸结合蛋白家族的巨大多样性，使用传统方法对 GTP 结合蛋白的表达水平、结构、活性和调控机制进行综合分析仍然具有挑战性。质谱 (MS) 仪器和数据采集方法的许多进步，以及样品制备中的各种富集方法，使 MS 成为全面表征各种 GTP 结合蛋白的活性和表达水平的强大工具。我们在此回顾了基于 MS 的技术应用的最新进展，以及通用和广泛使用的亲和富集方法，用于蛋白质组范围内的靶向捕获、鉴定和定量 GTP 结合蛋白。讨论了用于分析 GTP 结合蛋白的表达水平、活性、翻译后修饰和相互作用组的各种策略的工作原理、优点和局限性。可以设想，基于 MS 的蛋白质组学的未来应用将有助于更好地了解 GTP 结合蛋白在不同生物过程和人类疾病中的作用。© 2020 John Wiley & Sons Ltd. 质谱修订版。可以设想，基于 MS 的蛋白质组学的未来应用将有助于更好地了解 GTP 结合蛋白在不同生物过程和人类疾病中的作用。© 2020 John Wiley & Sons Ltd. 质谱修订版。可以设想，基于 MS 的蛋白质组学的未来应用将有助于更好地了解 GTP 结合蛋白在不同生物过程和人类疾病中的作用。© 2020 John Wiley & Sons Ltd. 质谱修订版。