title: Comparison of the methods for profiling glycoprotein glycans--HUPO Human Disease Glycomics/Proteome Initiative multi-institutional study.
abstract: Mass spectrometry (MS) of glycoproteins is an emerging field in proteomics, poised to meet the technical demand for elucidation of the structural complexity and functions of the oligosaccharide components of molecules. Considering the divergence of the mass spectrometric methods employed for oligosaccharide analysis in recent publications, it is necessary to establish technical standards and demonstrate capabilities. In the present study of the Human Proteome Organisation (HUPO) Human Disease Glycomics/Proteome Initiative (HGPI), the same samples of transferrin and immunoglobulin-G were analyzed for N-linked oligosaccharides and their relative abundances in 20 laboratories, and the chromatographic and mass spectrometric analysis results were evaluated. In general, matrix-assisted laser desorption/ionization (MALDI) time-of-flight MS of permethylated oligosaccharide mixtures carried out in six laboratories yielded good quantitation, and the results can be correlated to those of chromatography of reductive amination derivatives. For underivatized oligosaccharide alditols, graphitized carbon-liquid chromatography (LC)/electrospray ionization (ESI) MS detecting deprotonated molecules in the negative ion mode provided acceptable quantitation. The variance of the results among these three methods was small. Detailed analyses of tryptic glycopeptides employing either nano LC/ESI MS/MS or MALDI MS demonstrated excellent capability to determine site-specific or subclass-specific glycan profiles in these samples. Taking into account the variety of MS technologies and options for distinct protocols used in this study, the results of this multi-institutional study indicate that MS-based analysis appears as the efficient method for identification and quantitation of oligosaccharides in glycomic studies and endorse the power of MS for glycopeptide characterization with high sensitivity in proteomic programs.
authors.affiliation: Osaka Medical Center and Research Institute for Maternal and Child Health, 840 Murodo-cho, Izumi, Osaka 594-1101, Japan. waday@mch.pref.osaka.jp
authors.forename: Yoshinao
authors.initials: Y
authors.lastname: Wada
authors.forename: Parastoo
authors.initials: P
authors.lastname: Azadi
authors.forename: Catherine E
authors.initials: CE
authors.lastname: Costello
authors.forename: Anne
authors.initials: A
authors.lastname: Dell
authors.forename: Raymond A
authors.initials: RA
authors.lastname: Dwek
authors.forename: Hildegard
authors.initials: H
authors.lastname: Geyer
authors.forename: Rudolf
authors.initials: R
authors.lastname: Geyer
authors.forename: Kazuaki
authors.initials: K
authors.lastname: Kakehi
authors.forename: Niclas G
authors.initials: NG
authors.lastname: Karlsson
authors.forename: Koichi
authors.initials: K
authors.lastname: Kato
authors.forename: Nana
authors.initials: N
authors.lastname: Kawasaki
authors.forename: Kay-Hooi
authors.initials: KH
authors.lastname: Khoo
authors.forename: Soohyun
authors.initials: S
authors.lastname: Kim
authors.forename: Akihiro
authors.initials: A
authors.lastname: Kondo
authors.forename: Erika
authors.initials: E
authors.lastname: Lattova
authors.forename: Yehia
authors.initials: Y
authors.lastname: Mechref
authors.forename: Eiji
authors.initials: E
authors.lastname: Miyoshi
authors.forename: Kazuyuki
authors.initials: K
authors.lastname: Nakamura
authors.forename: Hisashi
authors.initials: H
authors.lastname: Narimatsu
authors.forename: Milos V
authors.initials: MV
authors.lastname: Novotny
authors.forename: Nicolle H
authors.initials: NH
authors.lastname: Packer
authors.forename: Hélène
authors.initials: H
authors.lastname: Perreault
authors.forename: Jasna
authors.initials: J
authors.lastname: Peter-Katalinic
authors.forename: Gottfried
authors.initials: G
authors.lastname: Pohlentz
authors.forename: Vernon N
authors.initials: VN
authors.lastname: Reinhold
authors.forename: Pauline M
authors.initials: PM
authors.lastname: Rudd
authors.forename: Akemi
authors.initials: A
authors.lastname: Suzuki
authors.forename: Naoyuki
authors.initials: N
authors.lastname: Taniguchi
mesh_terms: D002236:Carbohydrate Conformation; D020869:Gene Expression Profiling / Q000379:methods*; D030342:Genetic Diseases, Inborn / Q000235:genetics*; D006020:Glycopeptides / Q000737:chemistry; D006023:Glycoproteins / Q000737:chemistry / Q000235:genetics*; D006801:Humans; D013058:Mass Spectrometry; D008958:Models, Molecular; D009844:Oligosaccharides / Q000737:chemistry; D011134:Polysaccharides / Q000737:chemistry / Q000235:genetics*; D020543:Proteome; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D006020:Glycopeptides; D006023:Glycoproteins; D009844:Oligosaccharides; D011134:Polysaccharides; D020543:Proteome