Precursor ion scanning and sequencing of arginine-ADP-ribosylated peptide by mass spectrometry
Precursor ion scanning and sequencing of arginine-ADP-ribosylated peptide by mass spectrometry
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DOI：
10.1016/j.ab.2009.06.028
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发表时间：
2009-10-15
影响因子：
2.9
通讯作者：
Tsuchiya, Mikako
中科院分区：
文献类型：
--
作者：
Osago, Harumi;Yamada, Kazuo;Tsuchiya, Mikako
精氨酸(Arg)特异性的ADP核糖基化是蛋白质的翻译后修饰之一，被认为在真核生物中对靶蛋白功能的可逆调节起着重要作用。然而，目前还没有建立起生理靶蛋白。我们用四极串联质谱仪研究了ADP-R-Arg(ADP-R-Arg)和Arg-ADP-核糖化肽(ADP-R-Arg)的裂解模式，发现ADP-R-Arg特异性地裂解成N-(ADP-核糖基)-碳二亚胺(ADP-R-Carddiimide)和鸟氨酸。基于这种特殊的碎裂模式，我们用两步碰撞法成功地鉴定了Arg-ADP-核糖化肽的修饰位点和序列，表明ADP-R-碳二亚胺是Arg-ADP-核糖化肽前体离子扫描的良好标记离子。我们认为，前体离子扫描和ADP-R-碳二亚胺作为标记离子和两步碰撞相结合是寻找Arg-ADP-核糖化的生理靶蛋白的有效方法。(C)2009 Elsevier Inc.保留所有权利。
Arginine (Arg)-specific ADP-ribosylation is one of the posttranslational modifications of proteins and is thought to play an important role in reversibly regulating functions of the target proteins in eukaryotes. However, the physiological target protein has not been established. We examined the fragmentation pattern of both ADP-ribosyl-Arg (ADP-R-Arg) and Arg-ADP-ribosylated peptides by quadrupole tandem mass spectrometry and found a specific cleavage of ADP-R-Arg into N-(ADP-ribosyl)-carbodiimide (ADP-R-carbodiimide) and ornithine. Based on this specific fragmentation pattern, we successfully identified the modification site and sequence of Arg-ADP-ribosylated peptide using a two-step collision and showed that ADP-R-carbodiimide is an excellent marker ion for precursor ion scanning of Arg-ADP-ribosylated peptide. We propose that a combination of the precursor ion scanning with ADP-R-carbodiimide as a marker ion and two-step collision is useful in searching for physiological target proteins of Arg-ADP-ribosylation. (C) 2009 Elsevier Inc. All rights reserved.