Isotopically labeled crosslinking reagents: Resolution of mass degeneracy in the identification of crosslinked peptides
Isotopically labeled crosslinking reagents: Resolution of mass degeneracy in the identification of crosslinked peptides
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DOI：
10.1016/j.bmcl.2003.08.053
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发表时间：
2003-11-17
影响因子：
2.7
通讯作者：
Guy, RK
中科院分区：
文献类型：
--
作者：
Collins, CJ;Schilling, B;Guy, RK
三维质谱 (MS3D) 是一种新开发的蛋白质结构测定方法，涉及蛋白质的分子内化学交联、所得加合物的蛋白水解消化、通过质谱 (MS) 鉴定交联、使用理论质量列表进行峰分配以及通过距离几何方法计算减少结构交联。为了便于通过 MS 从交联蛋白的蛋白水解消化混合物中明确鉴定交联肽，我们在交联试剂中引入了双 O-18 同位素标记，为交联肽提供特征同位素模式。这些材料的质谱中存在相距 4 Da 的双峰，可以使用 MS/MS 数据的自动智能数据采集 (IDA) 轻松区分交联肽和修饰肽以及未交联肽。这应该允许该方法的自动化，以应用于整个可表达的蛋白质组。 (C) 2003 Elsevier Ltd. 保留所有权利。
Mass spectrometry in three dimensions (MS3D) is a newly developed method for the determination of protein structures involving intramolecular chemical crosslinking of proteins, proteolytic digestion of the resulting adducts, identification of crosslinks by mass spectrometry (MS), peak assignment using theoretical mass lists, and computational reduction of crosslinks to a structure by distance geometry methods. To facilitate the unambiguous identification of crosslinked peptides from proteolytic digestion mixtures of crosslinked proteins by MS, we introduced double O-18 isotopic labels into the crosslinking reagent to provide the crosslinked peptides with a characteristic isotope pattern. The presence of doublets separated by 4 Da in the mass spectra of these materials allowed ready discrimination between crosslinked and modified peptides, and uncrosslinked peptides using automated intelligent data acquisition (IDA) of MS/MS data. This should allow ready automation of the method for application to whole expressible proteomes. (C) 2003 Elsevier Ltd. All rights reserved.