基于等质量肽段末端标记策略的质谱鉴定新算法  

Novel Algorithm for Identification and Quantification of Proteins Based on Strategy of Isobaric Peptide Termini Labelling

作　　者：郑乃仁[1] 单亦初[2] 邓玉林[1] 张玉奎[1,2] 

机构地区：[1]北京理工大学生命学院,北京100081 [2]中国科学院大连化学物理研究所,大连116011

出　　处：《分析化学》2017年第10期1441-1447,共7页Chinese Journal of Analytical Chemistry

基　　金：国家重大科学仪器设备开发专项基金(No.2012YQ04014005)项目资助~~

摘　　要：等质量肽段末端标记(Isobaric peptide termini labeling,IPTL)是一种使用轻、重同位素分别对肽段的C端和N端进行等重标记的技术。在对使用这种标记技术得到的数据进行一级谱分析时,由于肽段的质量相同,不会增加样本的复杂性,而在处理二级谱的数据时,可利用成对的b、y离子进行分析。本研究利用IPTL方法得到的实验数据设计了一种新的打分算法:全部离子打分算法(All ions scoring algorithm,AISA)。AISA在对数据进行处理时,可以同时得到定性和定量信息。在Q-Exactive He La和Human-HCC-HL数据集上的蛋白定量覆盖率分别达到99%和100%。在Q-Exactive He La 2D RPLC数据集上,AISA算法鉴定到的PSM、唯一肽段和蛋白质分别比Morpheus高15%、26%和22%。在Human-HCC-HL数据集上,AISA算法鉴定到的PSM、唯一肽段和蛋白质分别比Morpheus高24%、39%和27%。在Q-Exactive He La和Human-HCC-HL数据集上蛋白质定量比值的平均值非常接近1,分别为1.18和0.90;在0.5~2.0区间内的定量比值分别为91%和94%。Isobaric peptide termini labeling（IPTL） is a technology which uses light and heavy isotopes to label C-terminus and N-terminus of peptides.As the masses of labeled peptides are equivalent,the complexity of sample is low when analyzing MS data produced by this technology.Besides,paired b and y ions are helpful while analyzing MS/MS data in this kind of experiments.On the basis of this,a novel scoring algorithm,all ions scoring algorithm（AISA）,has been designed for IPTL experiments.The information of quantification and qualification can be acquired at the same time using AISA.On Q-Exactive HeLa 2D RPLC dataset,peptide spectrum matches（PSMs）,distinct peptides and protein groups identified by AISA are 15%,26% and 22% higher than Morpheus.On human-HCC-HL dataset,PSMs,distinct peptides and protein groups identified by AISA are 24%,39% and 27% higher than Morpheus.Quantification ratio on Q-Exactive HeLa and humanHCC-HL datasets are 1.18 and 0.90,respectively,which are very close to 1.Besides,quantification ratios between 0.5 and 2.0 are 91% and 94%,respectively.

关 键 词：蛋白质组学 等重标记 蛋白质鉴定 

分 类 号：O629.73[理学—有机化学] O657.63[理学—化学]