主催: 日本ヒトプロテオーム機構
Glycosylation is the most abundant post translational modification of proteins. Analyses of glycopeptides of the N-liked type with MALD-TOF/TOF-MS provides for identification of the peptide as well as characterization of the glycan part and determination of glycosylation sites of glycoproteins.
2,5-Dihydroxybenzoic acid (DHB) is the matrix of choice for carbohydrate and glycopeptide analysis. We developed a sample preparation providing homogeneous sample morphology with low propensity to from sweet spots combined with a precisely defined sample dimension and location well suited for the unattended acquisition of spectra from DHB.
Automated LC-MALDI analysis of tryptic digests of glycoproteins permitted sensitive MS/MS analyses of glycopeptides on MALDI-TOF/TOF instrumentation: dedicated software a) provided automatically the list of glycopeptides, b) allowed the direct peptide identification via Mascot, and c) provided the modification site in the peptide and first insights into the glycostructure.
