A new crosslinker for mass spectrometric analysis of the quaternary structure of protein complexes
A new crosslinker for mass spectrometric analysis of the quaternary structure of protein complexes
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DOI：
10.1016/s1044-0305(00)00212-9
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发表时间：
2001-02-01
影响因子：
3.2
通讯作者：
de Jong, L
中科院分区：
文献类型：
--
作者：
Back, JW;Hartog, AF;de Jong, L
交联多肽的质谱结构分析是阐明多肽在蛋白质复合物中的空间排列的有力方法。我们的目标是开发双功能交联剂，在交联蛋白质复合物后，随后进行蛋白水解消化，产生交联肽，可以很容易地通过质谱法追踪。为此，我们合成了交联剂N-苄基亚氨基二乙酰氧基琥珀酰亚胺（BID），产生稳定的苄基阳离子标记离子后，低能量碰撞诱导解离（CID）串联质谱。用不同的BID交联肽制剂证明了在低能CID时标记离子的灵敏检测。使用BID，可以回收交联的和交联剂加合的肽，而无需在鉴定前纯化交联的肽。该交联剂的基本设计可以变化，以满足特定的交联需要，（C）2001 American Society for Mass Spectrometry。
Mass spectrometric structural analysis of crosslinked peptides is a powerful method to elucidate the spatial arrangement of polypeptides in protein complexes. Our aim is to develop bifunctional crosslinkers that, after crosslinking protein complexes followed by proteolytic digestion, give rise to crosslinked peptides that can be readily tracked down by mass spectrometry. To this end we synthesized the crosslinker N-benzyliminodiacetoyloxysuccinimid (BID), which yields stable benzyl cation marker ions upon low-energy collision-induced dissociation (CID) tandem mass spectrometry. Sensitive detection of the marker ion upon low-energy CID is demonstrated with different BID-crosslinked peptide preparations. With BID it becomes possible to retrieve crosslinked and crosslinker-adducted peptides, without the necessity of purifying crosslinked peptides prior to identification. The basic design of this crosslinker can be varied upon, in order to meet specific crosslinking needs, (C) 2001 American Society for Mass Spectrometry.