title: A strategy for identification of oligosaccharide structures using observational multistage mass spectral library.
abstract: Glycosylation is the most widespread posttranslational modification in eukaryotes; however, the role of oligosaccharides attached to proteins has been little studied because of the lack of a sensitive and easy analytical method for oligosaccharide structures. Recently, tandem mass spectrometric techniques have been revealing that oligosaccharides might have characteristic signal intensity profiles. We describe here a strategy for the rapid and accurate identification of the oligosaccharide structures on glycoproteins using only mass spectrometry. It is based on a comparison of the signal intensity profiles of multistage tandem mass (MSn) spectra between the analyte and a library of observational mass spectra acquired from structurally defined oligosaccharides prepared using glycosyltransferases. To smartly identify the oligosaccharides released from biological materials, a computer suggests which ion among the fragment ions in the MS/MS spectrum should yield the most informative MS3 spectrum to distinguish similar oligosaccharides. Using this strategy, we were able to identify the structure of N-linked oligosaccharides in immunoglobulin G as an example.
authors.affiliation: Research Center for Glycoscience, National Institute of Advanced Industrial Science and Technology, Tsukuba, Ibaraki 305-8568, Japan.
authors.forename: Akihiko
authors.initials: A
authors.lastname: Kameyama
authors.forename: Norihiro
authors.initials: N
authors.lastname: Kikuchi
authors.forename: Shuuichi
authors.initials: S
authors.lastname: Nakaya
authors.forename: Hiromi
authors.initials: H
authors.lastname: Ito
authors.forename: Takashi
authors.initials: T
authors.lastname: Sato
authors.forename: Toshihide
authors.initials: T
authors.lastname: Shikanai
authors.forename: Yoriko
authors.initials: Y
authors.lastname: Takahashi
authors.forename: Katsutoshi
authors.initials: K
authors.lastname: Takahashi
authors.forename: Hisashi
authors.initials: H
authors.lastname: Narimatsu
mesh_terms: D000595:Amino Acid Sequence; D016208:Databases, Factual; D006023:Glycoproteins / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D007074:Immunoglobulin G / Q000737:chemistry; D013058:Mass Spectrometry / Q000379:methods*; D008968:Molecular Conformation; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000032:analysis* / Q000737:chemistry*
chemical_list: D006023:Glycoproteins; D007074:Immunoglobulin G; D009844:Oligosaccharides