期刊
PROTEOMICS
卷 12, 期 22, 页码 3328-3337出版社
WILEY
DOI: 10.1002/pmic.201200150
关键词
Cell surface glycoprotein; CD-annotated protein; Glycopeptides; Glycoproteomics; Non-glycopeptides; Quantification
资金
- National Natural Sciences Foundation of China [20735004]
- Creative Research Group Project by NSFC [21021004]
- China State Key Basic Research Program Grant [2012CB910601, 2012CB910101]
- China High Technology Research Program Grant [2006AA02A309]
- National Key Special Program on Infection diseases [2012ZX10002009-011]
- Analytical Method Innovation Program of MOST [2009IM031800, 2010IM030500]
- Knowledge Innovation program of DICP
As the altered glycosylation expressions of cell surface proteins are associated with many diseases, glycoproteomics approach has been widely applied to characterization of surface glycosylation alteration. In general, the abundances of proteolytic glycopeptides derived from corresponding glycoproteins can be measured to determine the abundances of glycoproteins. However, this quantification strategy cannot distinguish whether the changes are results from changes of protein abundance or changes in glycosite occupancy. For the accurate and specific quantification of the cell surface glycosylation profile, we proposed a modified cell surface-capturing strategy where the glycopeptides were submitted to LC-MS/MS analysis directly for identification of glycoproteins and the non-glycopeptides were isotopically labelled for quantification of glycoproteins. This strategy was applied to comparatively analyze cell surface glycoproteins of two human cell lines, i.e. Chang Liver and HepG2 cells. Totally 341 glycoproteins were identified with 82.4% specificity for cell membrane proteins and 33 glycoproteins were quantified with significant expression change between the two cell lines. The differential expressions of two selected proteins (EMMPRIN and BCAM) were validated by Western blotting. This method enables specific and accurate analysis of the cell surface glycoproteins and may have broad application in the field of biomarker and drug target discovery.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据