规模化蛋白质生物质谱鉴定中肽段氨基端环化修饰现象论文.docVIP

　　规模化蛋白质生物质谱鉴定中肽段氨基端环化修饰现象论文 卢庄 赵丽艳 张养军 蔡耘 邓玉林 张玉奎 钱小红 【摘要】 对蛋白质样品制备中引入的氨基酸残基的一种现象——蛋白质酶切肽段氨基端的环化修饰现象的初步研究结果显示，很多以谷氨酰胺（Q）或氨乙酰化修饰的半胱氨酸（CAM_C）残基起始的肽段会发生氨基端的环化修饰，且修饰反应不完全，在同一样本中修饰与非修饰两种状态常同时存在，并且环化修饰后的肽段的反相色谱保留时间发生延迟。在数据库检索时添加环化修饰，可以提高蛋白质的鉴定成功率。本研究结果为大规模的蛋白质质谱数据解析提供了有价值的参考。 【关键词】 肽段氨基端，环化，规模化.freelass spectrometry has been developed for the largescale protein identification. The successful identification of protein in proteomic study is based on an effective match of MS data to the sequence in database. Because of the diversity and heterogeneity of protein modification, the experimental data obtained by mass spectrometry does not match the theoretical value sometimes, akes about 90 percent or more of the tandem mass spectra not be effectively identified. This has bee one of the most important technique issues to be resolved in current proteome research. The Nterminal cyclization of peptides, as one of a variety of modification introduced in sample preparation, has been preliminarily studied in this inal cyclization occurred at the most of the glutamine(Q) or carbamoylmethylcysteine(CAM_C) residues and the reaction is often inplete or partial, both types of peptides could often exist in its respective state at the same time, and the behavior of modified peptides in revered phase chromatography is also changed. The success rate of protein identification could be obviously improved if adding the Nterminal cyclization modification in the database searching. These results ass spectrometric data analysis of proteomic study. Keyinal of peptides, cyclization, largescale, protein identification 1 引 言 细胞或组织内存在的全部蛋白质的定性和定量分析是蛋白质组学研究的主要内容1。为了从细胞中鉴定目标蛋白质，候选蛋白质常用串联质谱（MS/MS）法2,3结合生物信息学的数据库检索进行分析。样品中的蛋白质通常先被胰蛋白酶消化成较小的肽段，然后经反相色谱分离进入质谱；肽段经离子化、碎裂后，产生用于鉴定的特征的MS/MS数据；通过在理论序列数据库中搜索MS/MS谱图来寻找最佳匹配的肽段，从而鉴定样本蛋白质。然而，在实际的蛋白质鉴定中，通常只有或少于10%的MS/MS谱图能够有效地鉴定出蛋白质，而大量的图谱都不能被解析4。 MS/MS数据没有得到可靠的鉴定结果，除了谱图质量差，还有一个原因就是匹配不好。很多碎片信息很丰富的MS/MS谱图在检索时的得分也很低，其原因是可能此肽段在数据库中不存在、或某些蛋白质本身存在，但人工处理过程中引入的各种氨基酸