title: Analysis of glycopeptides using lectin affinity chromatography with MALDI-TOF mass spectrometry.
abstract: Glycopeptides prepared from 1 nmol of a mixture of glycoproteins, transferrin, and ribonuclease B by lysylendopeptidase digestion were isolated by lectin and cellulose column chromatographies, and then they were analyzed by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry and MALDI-quadrupole ion trap (QIT)-TOF mass spectrometry which enables the performance of MS ( n ) analysis. The lectin affinity preparation of glycopeptides with Sambucus nigra agglutinin and concanavalin A provides the glycan structure outlines for the sialyl linkage and the core structure of N-glycans. Such structural estimation was confirmed by MALDI-TOF MS and MALDI-QIT-TOF MS/MS. Amino acid sequences and location of glycosylation sites were determined by MALDI-QIT-TOF MS/MS/MS. Taken together, the combination of lectin column chromatography, MALDI-TOF MS, and MALDI-QIT-TOF MS ( n ) provides an easy way for the structural estimation of glycans and the rapid analysis of glycoproteomics.
authors.affiliation: Research Team for Functional Genomics and Research Team for Molecular Biomarkers, Tokyo Metropolitan Institute of Gerontology, Foundation for Research on Aging and Promotion of Human Welfare, 35-2 Sakae-cho, Itabashi-ku, Tokyo 173-0015, Japan.
authors.forename: Kazutosi
authors.initials: K
authors.lastname: Kubota
authors.forename: Yuji
authors.initials: Y
authors.lastname: Sato
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Suzuki
authors.forename: Naoko
authors.initials: N
authors.lastname: Goto-Inoue
authors.forename: Tosifusa
authors.initials: T
authors.lastname: Toda
authors.forename: Minoru
authors.initials: M
authors.lastname: Suzuki
authors.forename: Shin-ichi
authors.initials: S
authors.lastname: Hisanaga
authors.forename: Akemi
authors.initials: A
authors.lastname: Suzuki
authors.forename: Tamao
authors.initials: T
authors.lastname: Endo
mesh_terms: D002846:Chromatography, Affinity / Q000379:methods*; D006020:Glycopeptides / Q000032:analysis* / Q000302:isolation & purification; D006801:Humans; D037102:Lectins / Q000737:chemistry*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
chemical_list: D006020:Glycopeptides; D037102:Lectins