Probing the exposure of the phosphate group in modified amino acids and peptides by ion-molecule reactions with triethoxyborane in Fourier transform ion cyclotron resonance mass spectrometry.
Probing the exposure of the phosphate group in modified amino acids and peptides by ion-molecule reactions with triethoxyborane in Fourier transform ion cyclotron resonance mass spectrometry.
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在傅里叶变换离子回旋共振质谱中，通过与三乙氧基硼烷的离子分子反应来探测修饰氨基酸和肽中磷酸基团的暴露。
DOI：
10.1002/rcm.6884
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发表时间：
2014
期刊：
影响因子：
--
通讯作者：
Lanucara F
中科院分区：
文献类型：
--
作者：
Lanucara F
理论基础磷酸基团和带电残基之间的分子内氢键在磷酸化肽的化学中起着至关重要的作用，驱动物质形成特定的构象并影响磷酸基团的暴露。这些相互作用的性质和程度可以通过测量磷酸基团对选定的底物在气相中的反应性进行研究。METHODSWe使用傅立叶变换离子回旋共振（FT-ICR）质谱（MS）进行系统的研究磷酸化的氨基酸和肽与三乙氧基硼烷（TEB）的气相离子反应性。通过电喷雾电离（ESI）产生感兴趣的离子，在FT-ICR质谱仪的池中分离，并使其与TEB的固定压力反应。反应的时间演变进行了监测和热速率常数的推导。离子产物的结构通过碰撞诱导解离（CID）串联质谱（MS/MS）进行了确认。研究发现，ATTSTEB与质子化磷酸化氨基酸和肽的磷酸根通过加成-消除途径反应。反应的动力学效率与试剂离子的电荷状态呈正相关，表明在反应过程中存在磷酸根基团对进入中性的电荷状态依赖性暴露。异构体磷酸化的肽，只有不同的修饰的丝氨酸残基的位置，表现出显着不同的动力学efficiency. CONCLUSIONSSThe能力的磷酸化的物种与TEB反应取决于在相应的气态离子的磷酸盐部分的访问的容易程度。测量这种反应的动力学效率可以代表探索生物分子中磷酸基团的可及性的有价值的工具。版权所有© 2014约翰威利父子有限公司.
RATIONALEIntramolecular hydrogen bonds between a phosphate group and charged residues play a crucial role in the chemistry of phosphorylated peptides, driving the species to specific conformations and affecting the exposure of the phosphate moiety. The nature and extent of these interactions can be investigated by measuring the reactivity of phosphate groups toward selected substrates in the gas phase.METHODSWe used Fourier Transform Ion Cyclotron Resonance (FT‐ICR) mass spectrometry (MS) to perform a systematic study on the gas‐phase ionic reactivity of phosphorylated amino acids and peptides with triethoxyborane (TEB). Ions of interest were generated by electrospray ionization (ESI), isolated in the cell of the FT‐ICR mass spectrometer, and allowed to react with a stationary pressure of TEB. The temporal evolution of the reaction was monitored and thermal rate constants were derived. The structure of the ionic products was confirmed by Collision‐Induced Dissociation (CID) tandem mass spectrometry (MS/MS).RESULTSTEB was found to react with the phosphate of protonated phosphorylated amino acids and peptides by an addition‐elimination pathway. The kinetic efficiency of the reaction showed a positive correlation with the charge state of the reagent ion, suggesting the existence of charge‐state‐dependent exposure of the phosphate groups towards the incoming neutral during the reaction. Isomeric phosphorylated peptides, only differing for the position of the modified serine residue, showed markedly different kinetic efficiencies.CONCLUSIONSThe ability of a phosphorylated species to react with TEB depends on the ease of access to the phosphate moiety in the corresponding gaseous ion. Measuring the kinetic efficiency of such reactions can represent a valuable tool to explore the accessibility of phosphate groups in biomolecules. Copyright © 2014 John Wiley & Sons, Ltd.
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