Quantitative analysis of protein expression using amine-specific isobaric tags in Escherichia coli cells expressing rhsA elements
Quantitative analysis of protein expression using amine-specific isobaric tags in Escherichia coli cells expressing rhsA elements
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DOI：
10.1002/pmic.200401231
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发表时间：
2005-06-01
期刊：
影响因子：
3.4
通讯作者：
Lee, KH
中科院分区：
文献类型：
--
作者：
Aggarwal, K;Choe, LH;Lee, KH
我们描述了使用胺特异性同量异序标签的蛋白质表达定量研究的影响rhsA元件在大肠杆菌中的过度表达。同量异位素标记策略的使用促进了鸟枪法蛋白质组学分析，并使多达四个样品的定量平行，报告离子系列的基础上，使用串联质谱（MS/MS）。使用液相色谱基质辅助激光解吸/电离方法，收集了23 139 MS/MS谱。对来自780种蛋白质的5063种肽进行了定量，包括几种丰度较低的蛋白质，如转录因子DnaB和DnaG。超过65%的蛋白质具有每个蛋白质至少两个高置信度肽匹配（p < 0.05）。此外，基于Grubb和Rosner检验的统计检验能够区分离群数据。删除离群数据对研究中代表的蛋白质的功能类别没有显著影响。
We describe the use of amine-specific isobaric tags for protein expression quantification to study the effect of rhsA element over-expression in Escherichia coli. The use of an isobaric tagging strategy facilitates a shotgun approach to proteomic analysis and enables quantitation of up to four samples in parallel, based on the reporter ion series using tandem mass spectrometry (MS/MS). Using a liquid chromatography matrix-assisted laser desorption/ionization approach, 23 139 MS/MS spectra were collected. Five thousand sixty-three peptides derived from 780 proteins were quantified including several lower abundance proteins, such as transcription factors, DnaB and DnaG. More than 65% of the proteins had at least two high confidence peptide matches per protein (p < 0.05). Further, a statistical test based on the Grubb's and Rosner's tests was able to discriminate outlier data. The removal of outlier data had no significant effect on the functional categories of proteins that were represented in the study.