title: PNGase F-mediated incorporation of (18)O into glycans for relative glycan quantitation.
abstract: PNGase F-catalyzed glycosylation site (18)O-labeling is a widely used method for glycoprotein quantitation owing to its efficiency and simplicity. However, PNGase F-catalyzed glycan (18)O-labeling, which offers advantages for glycomics, has not yet been developed. In this study, PNGase F-mediated incorporation of (18)O into glycans during the N-glycan release from glycoproteins by PNGase F was finally realized, named as PCGOL (PNGase F-catalyzed glycan (18)O-labeling), which offers a potential strategy for relative glycan quantitation. This new method showed good linearity and high reproducibility within at least 2 orders of magnitude in the dynamic range. Furthermore, PCGOL combined with our previously developed TOSIL method (tandem (18)O stable isotope labeling for N-glycoproteome quantitation) can be used for comprehensive N-glycosylation quantification, achieving simultaneous quantification of glycans, glycopeptides and glycoproteins in a single workflow, which was also used to analyze glycosylation changes in immunoglobulin G (IgG) associated with hepatocellular carcinoma in the present work.
authors.affiliation: Department of Chemistry, Fudan University, Shanghai 200433, P. R. China. pyyang@fudan.edu.cn.
authors.forename: Wei
authors.initials: W
authors.lastname: Zhang
authors.forename: Weiqian
authors.initials: W
authors.lastname: Cao
authors.forename: Jiangming
authors.initials: J
authors.lastname: Huang
authors.forename: Hong
authors.initials: H
authors.lastname: Wang
authors.forename: Ji
authors.initials: J
authors.lastname: Wang
authors.forename: Chen
authors.initials: C
authors.lastname: Xie
authors.forename: Pengyuan
authors.initials: P
authors.lastname: Yang
mesh_terms: D000595:Amino Acid Sequence; D002240:Carbohydrate Sequence; D006528:Carcinoma, Hepatocellular / Q000378:metabolism; D054794:Glycomics / Q000379:methods; D006020:Glycopeptides / Q000737:chemistry / Q000378:metabolism; D006023:Glycoproteins / Q000737:chemistry* / Q000378:metabolism; D006031:Glycosylation; D006801:Humans; D007074:Immunoglobulin G / Q000737:chemistry / Q000378:metabolism; D008113:Liver Neoplasms / Q000378:metabolism; D013058:Mass Spectrometry / Q000379:methods; D008969:Molecular Sequence Data; D010103:Oxygen Isotopes / Q000032:analysis / Q000378:metabolism*; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase / Q000378:metabolism*; D011134:Polysaccharides / Q000032:analysis* / Q000378:metabolism; D015203:Reproducibility of Results
chemical_list: D006020:Glycopeptides; D006023:Glycoproteins; D007074:Immunoglobulin G; D010103:Oxygen Isotopes; D011134:Polysaccharides; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase