Dissociation behavior of a bifunctional tempo-active ester reagent for peptide structure analysis by free radical initiated peptide sequencing (FRIPS) mass spectrometry.
Dissociation behavior of a bifunctional tempo-active ester reagent for peptide structure analysis by free radical initiated peptide sequencing (FRIPS) mass spectrometry.
复制标题
双功能时间活性酯试剂的解离行为，用于自由基引发肽测序 (FRIPS) 质谱分析肽结构
DOI：
10.1002/jms.3543
复制
发表时间：
2015
期刊：
影响因子：
--
通讯作者：
Schäfer M
中科院分区：
文献类型：
--
作者：
Ihling C;Falvo F;Kratochvil I;Sinz A;Schäfer M
我们合成了一种同双功能活性酯交联剂，含有连接到苄基（Bz）的 TEMPO（2,2,6,6-四甲基哌啶-1-氧基）部分，称为 TEMPO-Bz-连接体。设计这种新型交联剂的目的是促进通过自由基引发肽测序 (FRIPS) 对交联产物进行 MS 分析。 TEMPO-Bz-连接体与所有 20 种蛋白氨基酸以及模型肽发生反应，以详细了解其在碰撞激活时的断裂机制。这项原理验证研究的最终目标是评估 TEMPO-Bz 连接剂在化学交联研究中的潜力，以获取蛋白质的 3D 结构信息。我们的研究是由 TEMPO-Bz 试剂的中心 NO-C 键在碰撞激活时的不稳定性得到充分记录的。针对大量前体离子的电荷状态和氨基酸组成研究了该特定键的断裂，从而鉴定了两种不同的断裂途径。具有强碱性残基的分子离子能够保持电荷载体的位置，即质子或钠阳离子，并因此通过 TEMPO-Bz-连接体的 NO-C 键的均裂裂解而分解。这导致形成互补的开壳肽自由基阳离子，而在 TEMPO-Bz-连接体本身质子化的前体离子在碰撞激活时表现出电荷驱动的偶数电子产物离子的形成。 MS3产物离子实验提供了氨基酸序列信息并允许确定交联位点。我们的研究充分表征了 TEMPO-Bz-连接剂的 CID 行为，并展示了其潜力，但也揭示了其在基于选择性串联 MS 分析的基础上利用开壳肽离子的特殊功能进行化学交联应用的局限性。版权所有 © 2015 约翰·威利父子有限公司
We have synthesized a homobifunctional active ester cross‐linking reagent containing a TEMPO (2,2,6,6‐tetramethylpiperidine‐1‐oxy) moiety connected to a benzyl group (Bz), termed TEMPO‐Bz‐linker. The aim for designing this novel cross‐linker was to facilitate MS analysis of cross‐linked products by free radical initiated peptide sequencing (FRIPS). The TEMPO‐Bz‐linker was reacted with all 20 proteinogenic amino acids as well as with model peptides to gain detailed insights into its fragmentation mechanism upon collision activation. The final goal of thisproof‐of‐principlestudy was to evaluate the potential of the TEMPO‐Bz‐linker for chemical cross‐linking studies to derive 3D‐structure information of proteins. Our studies were motivated by the well documented instability of the central NO―C bond of TEMPO‐Bz reagents upon collision activation. The fragmentation of this specific bond was investigated in respect to charge states and amino acid composition of a large set of precursor ions resulting in the identification of two distinct fragmentation pathways. Molecular ions with highly basic residues are able to keep the charge carriers located, i.e. protons or sodium cations, and consequently decompose via a homolytic cleavage of the NO―C bond of the TEMPO‐Bz‐linker. This leads to the formation of complementary open‐shell peptide radical cations, while precursor ions that are protonated at the TEMPO‐Bz‐linker itself exhibit a charge‐driven formation of even‐electron product ions upon collision activation. MS3product ion experiments provided amino acid sequence information and allowed determining the cross‐linking site. Our study fully characterizes the CID behavior of the TEMPO‐Bz‐linker and demonstrates its potential, but also its limitations for chemical cross‐linking applications utilizing the special features of open‐shell peptide ions on the basis of selective tandem MS analysis. Copyright © 2015 John Wiley & Sons, Ltd.
登录
查看更多内容
单电荷 DEST 交联肽的独特断裂
影响因子：
3.2
作者：
Yi He;M. Lauber;J. Reilly
通讯作者：
J. Reilly
影响因子：
1.8
作者：
Falvo, Francesco;Fiebig, Lukas;Schaefer, Mathias
通讯作者：
Schaefer, Mathias
DOI：
10.1016/1044-0305(96)00019-0
发表时间：
1996-06-01
影响因子：
3.2
作者：
Cox, KA;Gaskell, SJ;Whiting, A
通讯作者：
Whiting, A
影响因子：
2.3
作者：
Lee, Minhee;Lee, Younjin;Oh, Han Bin
通讯作者：
Oh, Han Bin
影响因子：
4.2
作者：
Lee, Minhee;Kang, Minhyuck;Oh, Han Bin
通讯作者：
Oh, Han Bin