Studying O-linked protein glycosylations in human plasma.
Recent investigations have implicated aberrant glycosylations in various malignancies, including epithelial ovarian cancer (EOC). The protocol here identifies O-linked carbohydrate patterns in EOC plasma glycoproteins through chemical cleavage and purification of these glycans. Dialyzed plasma is subjected to reductive beta-elimination with alkaline borohydride to release O-linked oligosaccharides from glycoproteins. Enrichment of released glycans, as well as removal of peptide and other contaminants, is followed by carbohydrate pattern analysis with MALDI-FT-ICR-MS.
Williams
Taufika Islam
TI
W.M. Keck FT-ICR Mass Spectrometry Laboratory, Department of Chemistry, North Carolina State University, Raleigh, NC 27695, USA.
Saggese
Diana A
DA
Muddiman
David C
DC
D001798:Blood Proteins / Q000737:chemistry* / Q000378:metabolism; D001894:Borohydrides / Q000737:chemistry; D003956:Dialysis / Q000379:methods; D005260:Female; D006031:Glycosylation; D006595:Hexosamines / Q000032:analysis; D006601:Hexoses / Q000032:analysis; D006603:Hexuronic Acids / Q000032:analysis; D006801:Humans; D008875:Middle Aged; D010051:Ovarian Neoplasms / Q000097:blood*; D011134:Polysaccharides / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification; D011402:Pronase / Q000378:metabolism; D052616:Solid Phase Extraction / Q000379:methods; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods
D001798:Blood Proteins; D001894:Borohydrides; D006595:Hexosamines; D006601:Hexoses; D006603:Hexuronic Acids; D011134:Polysaccharides; C025364:sodium borohydride; D011402:Pronase