Utilizing Ion Mobility to Identify Isobaric Post-Translational Modifications: Resolving Acrolein and Propionyl Lysine Adducts by TIMS Mass Spectrometry.
Utilizing Ion Mobility to Identify Isobaric Post-Translational Modifications: Resolving Acrolein and Propionyl Lysine Adducts by TIMS Mass Spectrometry.
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DOI：
10.1007/s12127-018-0237-z
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发表时间：
2018-09
期刊：
影响因子：
--
通讯作者：
Fritz KS
中科院分区：
文献类型：
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作者：
Gomez JD;Ridgeway ME;Park MA;Fritz KS
蛋白质翻译后修饰为阐明因毒性暴露、代谢改变或疾病发病机制而导致的蛋白质功能改变的机制提供了关键的蛋白质组学细节。赖氨酸丙酰化是一种最近描述的修饰，它是由于丙酰辅酶A代谢和去乙酰化酶活性的改变而发生的。丙烯醛是一种通过外源性和内源性途径产生的有毒醛类，例如工业接触、吸入香烟烟雾和非酶促脂质过氧化。重要的是，由丙酰化和丙烯醛化产生的赖氨酸修饰可能是同质异位的——通过质谱无法区分——并且通过反相色谱无法分离。在此，我们介绍了一种新型的应用，即利用离子迁移谱（TIMS）来解析这种相互竞争的同质异位赖氨酸修饰。具体而言，我们使用离子迁移谱 - 飞行时间质谱仪（TIMS - TOF）的原型分析了一种小的合成肽的翻译后修饰产物。由于这些丙酰化和丙烯醛化肽的迁移率仅相差1%，因此需要高分辨率的迁移率分析来区分这两种物质。我们能够在离子迁移谱分析仪中实现足够高的分辨率（约170），从而轻松地分离这些同质异位体。
Protein post-translational modifications provide critical proteomic details towards elucidating mechanisms of altered protein function due to toxic exposure, altered metabolism, or disease pathogenesis. Lysine propionylation is a recently described modification that occurs due to metabolic alterations in propionyl-CoA metabolism and sirtuin depropionylase activity. Acrolein is a toxic aldehyde generated through exogenous and endogenous pathways, such as industrial exposure, cigarette smoke inhalation, and non-enzymatic lipid peroxidation. Importantly, lysine modifications arising from propionylation and acroleination can be isobaric – indistinguishable by mass spectrometry – and inseparable via reverse-phase chromatography. Here, we present the novel application of trapped ion mobility spectrometry (TIMS) to resolve such competing isobaric lysine modifications. Specifically, the PTM products of a small synthetic peptide were analyzed using a prototype TIMS – time-of-flight mass spectrometer (TIMS-TOF). In that the mobilities of these propionylated and acroleinated peptides differ by only 1%, a high-resolution mobility analysis is required to resolve the two. We were able to achieve more than sufficient resolution in the TIMS analyzer (~170), readily separating these isobars.