title: Large-scale identification of target proteins of a glycosyltransferase isozyme by Lectin-IGOT-LC/MS, an LC/MS-based glycoproteomic approach.
abstract: Model organisms containing deletion or mutation in a glycosyltransferase-gene exhibit various physiological abnormalities, suggesting that specific glycan motifs on certain proteins play important roles in vivo. Identification of the target proteins of glycosyltransferase isozymes is the key to understand the roles of glycans. Here, we demonstrated the proteome-scale identification of the target proteins specific for a glycosyltransferase isozyme, β1,4-galactosyltransferase-I (β4GalT-I). Although β4GalT-I is the most characterized glycosyltransferase, its distinctive contribution to β1,4-galactosylation has been hardly described so far. We identified a large number of candidates for the target proteins specific to β4GalT-I by comparative analysis of β4GalT-I-deleted and wild-type mice using the LC/MS-based technique with the isotope-coded glycosylation site-specific tagging (IGOT) of lectin-captured N-glycopeptides. Our approach to identify the target proteins in a proteome-scale offers common features and trends in the target proteins, which facilitate understanding of the mechanism that controls assembly of a particular glycan motif on specific proteins.
authors.affiliation: Research Center for Medical Glycoscience, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, Ibaraki, Japan.
authors.forename: Daisuke
authors.initials: D
authors.lastname: Sugahara
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Kaji
authors.forename: Kazushi
authors.initials: K
authors.lastname: Sugihara
authors.forename: Masahide
authors.initials: M
authors.lastname: Asano
authors.forename: Hisashi
authors.initials: H
authors.lastname: Narimatsu
mesh_terms: D000818:Animals; D002853:Chromatography, Liquid; D005260:Female; D005700:Galactosyltransferases / Q000235:genetics / Q000378:metabolism*; D017353:Gene Deletion; D006020:Glycopeptides / Q000032:analysis* / Q000378:metabolism; D006031:Glycosylation; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism; D037102:Lectins / Q000737:chemistry*; D008099:Liver / Q000201:enzymology*; D013058:Mass Spectrometry; D051379:Mice; D018345:Mice, Knockout; D010103:Oxygen Isotopes; D011134:Polysaccharides / Q000032:analysis* / Q000378:metabolism; D040901:Proteomics
chemical_list: D006020:Glycopeptides; D007527:Isoenzymes; D037102:Lectins; D010103:Oxygen Isotopes; D011134:Polysaccharides; D005700:Galactosyltransferases