A comparative study of glycoprotein concentration, glycoform profile and glycosylation site occupancy using isotope labeling and electrospray linear ion trap mass spectrometry

Analytica Chimica Acta
Chih-Yu LinGuor-Rong Her

Abstract

A strategy is presented for comparative analysis of glycoproteins in which the variation of protein concentration, variation of glycosylation site occupancy and variation of glycoform profile can be determined. A comparative study was performed using stable isotope labeling of glycopeptides and peptides by formaldehyde-H(2) and formaldehyde-D(2) and analysis by ESI-MS analysis. The relative intensity of the nonglycosylated peptide provided information about protein concentration variation. Variation of the glycoform profile was obtained by comparing the glycoform profile of d(0)- and d(4)-dimethyl labeled glycopeptides. By knowing the variation of protein concentration and the variation of glycoform profile, the variation of glycosylation site occupancy could be calculated. The utility of the proposed strategy was demonstrated with ribonuclease B with different protein concentrations, different levels of glycosylation site occupancy and different glycoform profiles.

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Citations

Feb 8, 2013·Molecular & Cellular Proteomics : MCP·Heather Desaire
Jul 11, 2014·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Masaki Kurogochi, Junko Amano
Sep 21, 2016·Philosophical Transactions. Series A, Mathematical, Physical, and Engineering Sciences·Jue-Liang Hsu, Shu-Hui Chen
Dec 8, 2015·Glycoconjugate Journal·Lucia F Zacchi, Benjamin L Schulz
Aug 16, 2019·Cells·Qiongyu LiCarlito B Lebrilla

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