Quaternary diamines as mass spectrometry cleavable crosslinkers for protein interactions.
Quaternary diamines as mass spectrometry cleavable crosslinkers for protein interactions.
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DOI：
10.1007/s13361-011-0288-4
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发表时间：
2012-02
影响因子：
3.2
通讯作者：
Andrews, Philip C.
中科院分区：
文献类型：
--
作者：
Clifford-Nunn, Billy;Showalter, H. D. Hollis;Andrews, Philip C.
绘制蛋白质相互作用及其动力学对于定义生理状态，建立有效的模型以了解细胞功能以及允许更有效地靶向新药至关重要。交联研究可以估计蛋白质的接近程度，确定蛋白质-蛋白质相互作用的位点，并有可能通过共价锁定它们以进行分析来提供动态相互作用的快照。几个主要的挑战与在质谱法中使用交联剂有关，特别是在复杂的混合物中。我们描述了MS-可裂解的交联剂含有环胺，其中解决了一些这些挑战的合成和表征。DC 4交联剂含有两个固有正电荷，其允许交联的肽通过碰撞诱导解离（CID）或源内衰变片段化为其组分肽。初始片段化事件导致正电荷两侧的裂解，因此交联肽被鉴定为通过限定质量分离的离子对。然后，组分肽的结构可以通过MS 3稳健地确定，因为它们的片段化产物重排以产生移动的质子。DC 4交联剂对储存稳定、高反应性、高可溶性（1 M溶液）、对CID非常不稳定，并且MS 3导致生产性主链断裂。
Mapping protein interactions and their dynamics is crucial to defining physiologic states, building effective models for understanding cell function, and to allow more effective targeting of new drugs. Crosslinking studies can estimate the proximity of proteins, determine sites of protein–protein interactions, and have the potential to provide a snapshot of dynamic interactions by covalently locking them in place for analysis. Several major challenges are associated with the use of crosslinkers in mass spectrometry, particularly in complex mixtures. We describe the synthesis and characterization of a MS-cleavable crosslinker containing cyclic amines, which address some of these challenges. The DC4 crosslinker contains two intrinsic positive charges, which allow crosslinked peptides to fragment into their component peptides by collision-induced dissociation (CID) or in-source decay. Initial fragmentation events result in cleavage on either side of the positive charges so crosslinked peptides are identified as pairs of ions separated by defined masses. The structures of the component peptides can then be robustly determined by MS3 because their fragmentation products rearrange to generate a mobile proton. The DC4 crosslinking reagent is stable to storage, highly reactive, highly soluble (1 M solutions), quite labile to CID, and MS3 results in productive backbone fragmentation.
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影响因子：
2
作者：
Krauth, Fabian;Ihling, Christian H.;Sinz, Andrea
通讯作者：
Sinz, Andrea
影响因子：
7.4
作者：
King, Gordon J.;Jones, Alun;Ross, Ian L.
通讯作者：
Ross, Ian L.
影响因子：
2
作者：
Kang, Sebyung;Mou, Liyuan;Lanman, Jason;Velu, Sadanandan;Brouillette, Wayne J.;Prevelige, Peter E., Jr.
通讯作者：
Prevelige, Peter E., Jr.
影响因子：
2.7
作者：
Collins, CJ;Schilling, B;Guy, RK
通讯作者：
Guy, RK
DOI：
10.1016/j.jasms.2008.11.009
发表时间：
2009-04-01
影响因子：
3.2
作者：
Iglesias, Amadeu H.;Santos, Luiz F. A.;Gozzo, Fabio C.
通讯作者：
Gozzo, Fabio C.