An automated method for determining glycosylation and site diversity in glycoproteins

CARB 18

Carlito B Lebrilla1, Hyunjoo An2, John Tellinghast3, and David Rocke3. (1) Department of Chemistry, University of California, Davis, One shields Avenue, Davis, CA 95616, (2) Department of Chemistry, University of California, One Shields Ave, Davis, CA 95616, (3) Center for Image Processing and Integrated Computing, University of California, One Shields Ave, Davis, CA 95616
The determination of glycosylation sites and oligosaccharide heterogeneity is key toward understanding the biological roles of glycoproteins. There are no methods that can reliably determine the site of glycosylation and the glycan heterogeneity at specific sites. We have developed a procedure for the simultaneous determination of N- and O-glycosylation sites and oligosaccharide heterogeneity in glycoproteins. The method is based on a combination of non-specific proteolysis, deglycosylation, and high mass accuracy mass spectrometry analysis. The glycoprotein was proteolytically degraded into glycopeptide fragments. The exact peptide mass was calculated by subtracting the observed glycan mass from the observed glycopeptide mass. The amino acid sequence of a matched peptide mass was identified from the protein database. A computer program, Glyco X, was developed in MATLab and only requires the entry of the exact masses of the glycopeptide and glycan spectra in the form of an m/z table from the instrument data system.
 

Chemical Glycobiology Symposium
9:00 AM-12:20 PM, Monday, 11 September 2006 Hilton San Francisco -- Yosemite C, Oral

Division of Carbohydrate Chemistry

The 232nd ACS National Meeting, San Francisco, CA, September 10-14, 2006