An MRM-based workflow for absolute quantitation of lysine-acetylated metabolic enzymes in mouse liver.
An MRM-based workflow for absolute quantitation of lysine-acetylated metabolic enzymes in mouse liver.
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DOI：
10.1039/c5an01832c
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发表时间：
2015-11
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影响因子：
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通讯作者：
Leilei Xu;F. Wang;Ying-Hua Xu;Yi Wang;Cuiping Zhang;Xue Qin;Hongxiu Yu;Pengyuan Yang
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文献类型：
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作者：
Leilei Xu;F. Wang;Ying-Hua Xu;Yi Wang;Cuiping Zhang;Xue Qin;Hongxiu Yu;Pengyuan Yang
作为一种关键的翻译后修饰机制，蛋白质乙酰化在调节和/或协调细胞代谢方面起着至关重要的作用。乙酰化是酶中普遍存在的一种修饰过程。蛋白质的乙酰化修饰发生在亚化学计量的量，因此从这些乙酰化位点提取有生物学意义的信息需要一种适应的、敏感的、特异的和健壮的定量方法。在这项工作中，我们将免疫分析和多反应监测-质谱仪(MRM-MS)技术相结合，建立了乙酰化修饰的绝对定量方法。利用这种混合方法，我们对代谢酶的乙酰化水平进行了量化，从而可以揭示所研究的酶的调控机制。这一定量工作流程的发展是推进我们对蛋白质乙酰化在生理学和病理生理学中的调节作用的认识和理解的关键一步。
As a key post-translational modification mechanism, protein acetylation plays critical roles in regulating and/or coordinating cell metabolism. Acetylation is a prevalent modification process in enzymes. Protein acetylation modification occurs in sub-stoichiometric amounts; therefore extracting biologically meaningful information from these acetylation sites requires an adaptable, sensitive, specific, and robust method for their quantification. In this work, we combine immunoassays and multiple reaction monitoring-mass spectrometry (MRM-MS) technology to develop an absolute quantification for acetylation modification. With this hybrid method, we quantified the acetylation level of metabolic enzymes, which could demonstrate the regulatory mechanisms of the studied enzymes. The development of this quantitative workflow is a pivotal step for advancing our knowledge and understanding of the regulatory effects of protein acetylation in physiology and pathophysiology.