Comparison of the membrane proteome of virulent Mycobacterium tuberculosis and the attenuated Mycobacterium bovis BCG vaccine strain by label-free quantitative proteomics.
Comparison of the membrane proteome of virulent Mycobacterium tuberculosis and the attenuated Mycobacterium bovis BCG vaccine strain by label-free quantitative proteomics.
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DOI：
10.1021/pr400334k
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发表时间：
2013-12-06
影响因子：
4.4
通讯作者：
Chen X
中科院分区：
文献类型：
--
作者：
Gunawardena HP;Feltcher ME;Wrobel JA;Gu S;Braunstein M;Chen X
结核分枝杆菌（MTB）膜富含抗原，这些抗原是诊断和开发新疫苗的潜在靶点。为了更好地了解结核分枝杆菌毒力的机制和确定新的治疗干预的目标，我们研究了有毒的结核分枝杆菌之间的膜蛋白质组的差异组成。结核分枝杆菌H37 Rv（MTB）和牛分枝杆菌BCG疫苗株。为了比较膜蛋白质组，我们使用LC-MS/MS分析结合无标记定量（LFQ）蛋白质组学，利用从MS 1特征的m/z和保留时间对齐获得的肽的提取离子色谱图（XIC）的曲线下面积（AUC）。通过这种方法，我们获得了2，203个高置信度的膜相关蛋白的相对丰度比。在这些蛋白质中，294种在MTB和BCG膜级分之间的相对丰度上显示出至少2倍的统计学显著差异。我们的比较分析检测到与MTB和BCG之间的已知基因组差异区域相关的几种蛋白质不存在，这验证了我们方法的准确性。为了进一步支持我们的无标记定量数据，我们通过免疫印迹验证了选择的蛋白质差异。据我们所知，我们已经产生了第一个全面的和高覆盖率的比较膜蛋白质组变化之间的毒性MTB和其减毒的相对BCG，这有助于阐明的MTB病原体的内在毒力的蛋白质组学基础。
The Mycobacterium tuberculosis (MTB) membrane is rich in antigens that are potential targets for diagnostics and the development of new vaccines. To better understand the mechanisms underlying MTB virulence and identify new targets for therapeutic intervention we investigated the differential composition of membrane proteomes between virulent M. tuberculosis H37Rv (MTB) and the Mycobacterium bovis BCG vaccine strain. To compare the membrane proteomes, we used LC-MS/MS analysis in combination with label-free quantitative (LFQ) proteomics, utilizing the area-under-curve (AUC) of the extracted ion chromatograms (XIC) of peptides obtained from m/z and retention time alignment of MS1 features. With this approach, we obtained relative abundance ratios for 2,203 identified membrane-associated proteins in high confidence. Of these proteins, 294 showed statistically significant differences of at least 2 fold, in relative abundance between MTB and BCG membrane fractions. Our comparative analysis detected several proteins associated with known genomic regions of difference between MTB and BCG as being absent, which validated the accuracy of our approach. In further support of our label-free quantitative data, we verified select protein differences by immunoblotting. To our knowledge we have generated the first comprehensive and high coverage profile of comparative membrane proteome changes between virulent MTB and its attenuated relative BCG, which helps elucidate the proteomic basis of the intrinsic virulence of the MTB pathogen.
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DOI：
10.1074/jbc.m809088200
发表时间：
2009-05-08
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Hatzios SK;Schelle MW;Holsclaw CM;Behrens CR;Botyanszki Z;Lin FL;Carlson BL;Kumar P;Leary JA;Bertozzi CR
通讯作者：
Bertozzi CR
影响因子：
3.6
作者：
Dubey, VS;Sirakova, TD;Kolattukudy, PE
通讯作者：
Kolattukudy, PE
影响因子：
7
作者：
Gu, S;Chen, J;Chen, X
通讯作者：
Chen, X
影响因子：
56.9
作者：
Brightbill, HD;Libraty, DH;Modlin, RL
通讯作者：
Modlin, RL
影响因子：
3.6
作者：
Charlet, D;Mostowy, S;Behr, MA
通讯作者：
Behr, MA