Novel Concepts of MS-Cleavable Cross-linkers for Improved Peptide Structure Analysis
Novel Concepts of MS-Cleavable Cross-linkers for Improved Peptide Structure Analysis
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用于改进肽结构分析的 MS 可裂解交联剂的新概念
DOI：
10.1007/s13361-017-1712-1
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发表时间：
2022
影响因子：
3.2
通讯作者：
Sinz A
中科院分区：
文献类型：
--
作者：
Hage C;Falvo F;Schäfer M;Sinz A
化学交联/质谱（MS）方法作为研究蛋白质构象和破译蛋白质相互作用网络的替代方法越来越重要。这项研究是我们不断努力开发创新的交联原理的一部分，以方便和有效地分配交联产品。我们评估了两种同源双功能、胺反应性和质谱可切割的交联剂对交联产物自动分析的潜力。我们介绍了溴苯脲（BrPU）接头，它具有独特的结构，在碰撞激活时产生独特的碎片模式。此外，BrPU提供了特征的溴同位素模式和质量缺陷的所有交联剂修饰碎片。我们将BrPU连接子的断裂行为与我们之前描述的ms可切割的TEMPO-Bz连接子（由连接到苄基的2,2,6,6-四甲基哌啶-1-氧片段组成）进行了比较，后者被开发用于进行自由基启动的肽测序。在负电喷雾电离模式下，利用Orbitrap聚变质谱仪对这两种交联剂的碰撞激活实验（碰撞诱导解离和高能碰撞诱导解离）进行了比较。正如先前研究中假设的那样，交联n端天冬氨酸残基的存在似乎是交联产物中完整肽丢失的先决条件。由于BrPU连接器结合了特征质量位移和同位素特征，因此与TEMPO-Bz连接器相比，它提供了更有利的交联产物自动分配组合。
The chemical cross-linking/mass spectrometry (MS) approach is gaining increasing importance as an alternative method for studying protein conformation and for deciphering protein interaction networks. This study is part of our ongoing efforts to develop innovative cross-linking principles for a facile and efficient assignment of cross-linked products. We evaluate two homobifunctional, amine-reactive, and MS-cleavable cross-linkers regarding their potential for automated analysis of cross-linked products. We introduce the bromine phenylurea (BrPU) linker that possesses a unique structure yielding a distinctive fragmentation pattern on collisional activation. Moreover, BrPU delivers the characteristic bromine isotope pattern and mass defect for all cross-linker-decorated fragments. We compare the fragmentation behavior of the BrPU linker with that of our previously described MS-cleavable TEMPO-Bz linker (which consists of a 2,2,6,6-tetramethylpiperidine-1-oxy moiety connected to a benzyl group) that was developed to perform free-radical-initiated peptide sequencing. Comparative collisional activation experiments (collision-induced dissociation and higher-energy collision-induced dissociation) with both cross-linkers were conducted in negative electrospray ionization mode with an Orbitrap Fusion mass spectrometer using five model peptides. As hypothesized in a previous study, the presence of a cross-linked N-terminal aspartic acid residue seems to be the prerequisite for the loss of an intact peptide from the cross-linked products. As the BrPU linker combines a characteristic mass shift with an isotope signature, it presents a more favorable combination for automated assignment of cross-linked products compared with the TEMPO-Bz linker.
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影响因子：
7.4
作者：
Giese, Sven H.;Belsom, Adam;Rappsilber, Juri
通讯作者：
Rappsilber, Juri
利用酪氨酸特异性溴化同时鉴定酪氨酸磷酸化和硫酸化位点
影响因子：
3.2
作者：
Jong;Si;Hie
通讯作者：
Hie
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DOI：
10.1039/c5an02171e
发表时间：
2016
期刊：
The Analyst
影响因子：
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作者：
Y. Qi;D. Volmer
通讯作者：
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DOI：
10.1002/jms.3539
发表时间：
2015
期刊：
Journal of mass spectrometry : JMS
影响因子：
--
作者：
Jungjoo Nam;Hyuksu Kwon;Inae Jang;Aeran Jeon;Jingyu Moon;Sun Young Lee;Dukjin Kang;S. Han;Bongjin Moon;H. Oh
通讯作者：
H. Oh
影响因子：
4.4
作者：
Yang, Li;Zheng, Chunxiang;Weisbrod, Chad R.;Tang, Xiaoting;Munske, Gerhard R.;Hoopmann, Michael R.;Eng, Jimmy K.;Bruce, James E.
通讯作者：
Bruce, James E.