Chen Sharon S, Deutsch Eric W, Yi Eugene C, Li Xiao-jun, Goodlett David R, Aebersold Ruedi
University of Washington, Department of Bioengineering, Seattle, Washington 98105, USA.
J Proteome Res. 2005 Nov-Dec;4(6):2174-84. doi: 10.1021/pr050251c.
We present a method for peptide and protein identification based on LC-MS profiling. The method identified peptides at high-throughput without expending the sequencing time necessary for CID spectra based identification. The measurable peptide properties of mass and liquid chromatographic elution conditions are used to characterize and differentiate peptide features, and these peptide features are matched to a reference database from previously acquired and archived LC-MS/MS experiments to generate sequence assignments. The matches are scored according to the probability of an overlap between the peptide feature and the database peptides resulting in a ranked list of possible peptide sequences for each peptide submitted. This method resulted in 6 times more peptide sequence identifications from a single LC-MS analysis of yeast than from shotgun peptide sequencing using LC-MS/MS.
我们提出了一种基于液相色谱-质谱(LC-MS)分析的肽和蛋白质鉴定方法。该方法能够高通量鉴定肽段,而无需花费基于碰撞诱导解离(CID)光谱鉴定所需的测序时间。利用可测量的肽段质量和液相色谱洗脱条件等特性来表征和区分肽段特征,并将这些肽段特征与来自先前获取和存档的LC-MS/MS实验的参考数据库进行匹配,以生成序列分配。根据肽段特征与数据库肽段之间重叠的概率对匹配结果进行评分,从而为每个提交的肽段生成一份可能的肽段序列排名列表。与使用LC-MS/MS进行鸟枪法肽段测序相比,该方法对酵母进行单次LC-MS分析所鉴定出的肽段序列数量多出6倍。