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Gel-free mass spectrometry-based high throughput proteomics: Tools for studying biological response of proteins and proteomes

机译:基于无凝胶质谱的高通量蛋白质组学:研究蛋白质和蛋白质组生物学反应的工具

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摘要

Revolutionary advances in biological mass spectrometry (MS) have provided a basic tool to make possible comprehensive proteomic analysis. Traditionally, two-dimensional gel electrophoresis has been used as a separation method coupled with MS to facilitate analysis of complex protein mixtures. Despite the utility of this method, the many challenges of comprehensive proteomic analysis has motivated the development of gel-free MS-based strategies to obtain information not accessible using two-dimensional gel separations. These advanced strategies have enabled researchers to dig deeper into complex proteomes, gaining insights into the composition, quantitative response, covalent modifications and macromolecular interactions of proteins that collectively drive cellular function. This review describes the current state of gel-free, high throughput proteomic strategies using MS, including (i) the separation approaches commonly used for complex mixture analysis; (ii) strategies for large-scale quantitative analysis; (iii) analysis of post-translational modifications; and (iv) recent advances and future directions. The use of these strategies to make new discoveries at the proteome level into the effects of disease or other cellular perturbations is discussed in a variety of contexts, providing information on the potential of these tools in electromagnetic field research.
机译:生物质谱学(MS)的革命性进步为实现全面的蛋白质组学分析提供了基本工具。传统上,二维凝胶电泳已被用作与MS结合的分离方法,以促进复杂蛋白质混合物的分析。尽管该方法很有用,但是综合蛋白质组学分析的许多挑战促使开发无凝胶的基于MS的策略,以获取使用二维凝胶分离无法获得的信息。这些先进的策略使研究人员能够更深入地研究复杂的蛋白质组,深入了解共同驱动细胞功能的蛋白质的组成,定量反应,共价修饰和高分子相互作用。这篇综述描述了使用MS的无凝胶,高通量蛋白质组学策略的当前状态,包括:(i)复杂混合物分析中常用的分离方法; (ii)大规模定量分析的策略; (iii)分析翻译后修饰; (iv)最新进展和未来方向。在各种情况下讨论了使用这些策略在蛋白质组水平上发现新疾病或其他细胞扰动的影响的新发现,从而提供了有关这些工具在电磁场研究中的潜力的信息。

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