Structural analysis of an N-glycan with "beta1-4 bisecting branch" from human serum IgG by negative-ion MSn spectral matching and exoglycosidase digestion.
A novel N-linked oligosaccharide (N-glycan) with "beta1-4 bisecting branch (galactose beta1-4 bisecting N-acetylglucosamine)" was found in human serum IgG. Its structure was efficiently analyzed by using beta-galactosidase digestion, a MSn spectral library database, and negative-ion MS2 spectral matching. For confirmation, the novel N-glycan was synthesized by using an expected standard N-glycan (acceptor), UDP-galactose (donor), and beta1-4 galactosyltransferase. This work also demonstrates that the MSn spectral library database, in particular, negative-ion MS2 spectral matching, can efficiently reduce the number of specific, sequential exoglycosidase digestions required and is useful for rapid structural analysis of unknown glycans not in the database.
Takegawa
Yasuhiro
Y
Division of Biological Sciences, Graduate School of Science, Hokkaido University, Sapporo 001-0021, Japan.
Deguchi
Kisaburo
K
Nakagawa
Hiroaki
H
Nishimura
Shin-Ichiro
S
D000818:Animals; D002417:Cattle; D005690:Galactose / Q000737:chemistry / Q000378:metabolism; D005700:Galactosyltransferases / Q000378:metabolism; D006026:Glycoside Hydrolases / Q000378:metabolism*; D006801:Humans; D007074:Immunoglobulin G / Q000032:analysis* / Q000097:blood / Q000737:chemistry*; D007477:Ions / Q000737:chemistry*; D013058:Mass Spectrometry; D011134:Polysaccharides / Q000032:analysis* / Q000737:chemistry* / Q000378:metabolism; D011487:Protein Conformation; D013997:Time Factors
D007074:Immunoglobulin G; D007477:Ions; D011134:Polysaccharides; D005700:Galactosyltransferases; D006026:Glycoside Hydrolases; D005690:Galactose