Proteome-wide Characterization of N-Glycosylation Events by Diagonal Chromatography
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- 作者：Bart Ghesquiè ; re ; Jozef Van Damme ; Lennart Martens ; Joë ; l Vandekerckhove ; Kris Gevaert
- 刊名：Journal of Proteome Research
- 出版年：2006
- 出版时间：September 2006
- 年：2006
- 卷：5
- 期：9
- 页码：2438 - 2447
- 全文大小：229K
- 年卷期：v.5,no.9(September 2006)
- ISSN：1535-3907
文摘
A procedure to map N-glycosylation sites is presented here. It can be applied to purified proteins aswell as to highly complex mixtures. The method exploits deglycosylation by PNGase F in a diagonal,reverse-phase chromatographic setup. When applied to 10 
L of mouse serum, affinity-depleted for itsthree most abundant components, 117 known or predicted sites were mapped in addition to 10 novelsites. Several sites were detected on soluble membrane or receptor components. Our methodfurthermore senses the nature of glycan structures and can detect differential glycosylation on a givensite. These properties-high sensitivity and dependence on glycan imprinting-can be exploited forglycan-biomarker analysis.
L of mouse serum, affinity-depleted for itsthree most abundant components, 117 known or predicted sites were mapped in addition to 10 novelsites. Several sites were detected on soluble membrane or receptor components. Our methodfurthermore senses the nature of glycan structures and can detect differential glycosylation on a givensite. These properties-high sensitivity and dependence on glycan imprinting-can be exploited forglycan-biomarker analysis.