A strategy for identification of oligosaccharide structures using observational multistage mass spectral library.
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.
Kameyama
Akihiko
A
Research Center for Glycoscience, National Institute of Advanced Industrial Science and Technology, Tsukuba, Ibaraki 305-8568, Japan.
Kikuchi
Norihiro
N
Nakaya
Shuuichi
S
Ito
Hiromi
H
Sato
Takashi
T
Shikanai
Toshihide
T
Takahashi
Yoriko
Y
Takahashi
Katsutoshi
K
Narimatsu
Hisashi
H
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*
D006023:Glycoproteins; D007074:Immunoglobulin G; D009844:Oligosaccharides