Analysis of glycopeptides using lectin affinity chromatography with MALDI-TOF mass spectrometry.
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.
Kubota
Kazutosi
K
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.
Sato
Yuji
Y
Suzuki
Yusuke
Y
Goto-Inoue
Naoko
N
Toda
Tosifusa
T
Suzuki
Minoru
M
Hisanaga
Shin-ichi
S
Suzuki
Akemi
A
Endo
Tamao
T
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*
D006020:Glycopeptides; D037102:Lectins