Fragmentation behavior of a thiourea-based reagent for protein structure analysis by collision-induced dissociative chemical cross-linking.

Fragmentation behavior of a thiourea-based reagent for protein structure analysis by collision-induced dissociative chemical cross-linking.
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通过碰撞诱导解离化学交联进行蛋白质结构分析的硫脲基试剂的断裂行为

DOI:
10.1002/jms.1775
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发表时间:
2010
期刊:
Journal of mass spectrometry : JMS
影响因子:
--
通讯作者:
Schäfer
Schäfer
中科院分区:
--
文献类型:
--
作者:
Müller;Dreiocker;Ihling;Schäfer

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描述了一种专为碰撞诱导解离(CID)MS/MS实验设计的新型硫脲基交联剂分子的裂解行为。这种交联剂的开发是我们正在努力合成新型试剂的一部分,这些试剂在串联质谱法期间产生特征碎片离子或指示性恒定中性损失(CNL),从而可以对交联产物进行选择性和灵敏度分析。用于化学交联的新衍生试剂仅包含硫脲部分,其两侧是两个胺反应性N-羟基琥珀酰亚胺(NHS)酯部分,用于与蛋白质中的赖氨酸或游离N-末端反应。新试剂提供了简单的合成途径和两端长度或官能度的简单结构变化。硫脲部分具有特定的CID裂解能力-特征CNL为85 u-确保通过电喷雾电离(ESI)和基质辅助激光解吸/电离(MALDI)串联质谱法可靠检测衍生肽,因此具有化学交联研究的多功能适用性。对所提出的硫脲基试剂的CID行为的详细检查表明,试剂的轻微结构变化对于确保其全面有效地应用于蛋白质的化学交联是必要的。版权所有© 2010约翰威利父子有限公司.
The fragmentation behavior of a novel thiourea‐based cross‐linker molecule specifically designed for collision‐induced dissociation (CID) MS/MS experiments is described. The development of this cross‐linker is part of our ongoing efforts to synthesize novel reagents, which create either characteristic fragment ions or indicative constant neutral losses (CNLs) during tandem mass spectrometry allowing a selective and sensitive analysis of cross‐linked products. The new derivatizing reagent for chemical cross‐linking solely contains a thiourea moiety that is flanked by two amine‐reactiveN‐hydroxy succinimide (NHS) ester moieties for reaction with lysines or freeN‐termini in proteins. The new reagent offers simple synthetic access and easy structural variation of either length or functionalities at both ends. The thiourea moiety exhibits specifically tailored CID fragmentation capabilities—a characteristic CNL of 85 u—ensuring a reliable detection of derivatized peptides by both electrospray ionization (ESI) and matrix‐assisted laser desorption/ionization (MALDI) tandem mass spectrometry and as such possesses a versatile applicability for chemical cross‐linking studies. A detailed examination of the CID behavior of the presented thiourea‐based reagent reveals that slight structural variations of the reagent will be necessary to ensure its comprehensive and efficient application for chemical cross‐linking of proteins. Copyright © 2010 John Wiley & Sons, Ltd.
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