Differential Isotope Labeling of Glycopeptides for Accurate Determination of Differences in Site-Specific Glycosylation
Jazyk angličtina Země Spojené státy americké Médium print-electronic
Typ dokumentu časopisecké články, práce podpořená grantem
- Klíčová slova
- IgG, LC−ESI−MS, MALDI MS, glycopeptides, stable isotope labeling,
- MeSH
- anhydridy kyseliny jantarové chemie MeSH
- glykopeptidy chemie MeSH
- glykosylace MeSH
- hmotnostní spektrometrie s elektrosprejovou ionizací MeSH
- imunoglobulin G chemie MeSH
- izotopové značení MeSH
- lidé MeSH
- posttranslační úpravy proteinů * MeSH
- proteomika metody MeSH
- spektrometrie hmotnostní - ionizace laserem za účasti matrice MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- anhydridy kyseliny jantarové MeSH
- glykopeptidy MeSH
- imunoglobulin G MeSH
- succinic anhydride MeSH Prohlížeč
We introduce a stable isotope labeling approach for glycopeptides that allows a specific glycosylation site in a protein to be quantitatively evaluated using mass spectrometry. Succinic anhydride is used to specifically label primary amino groups of the peptide portion of the glycopeptides. The heavy form (D4(13)C4) provides an 8 Da mass increment over the light natural form (H4(12)C4), allowing simultaneous analysis and direct comparison of two glycopeptide profiles in a single MS scan. We have optimized a protocol for an in-solution trypsin digestion, a one-pot labeling procedure, and a post-labeling solid-phase extraction to obtain purified and labeled glycopeptides. We provide the first demonstration of this approach by comparing IgG1 Fc glycopeptides from polyclonal IgG samples with respect to their galactosylation and sialylation patterns using MALDI MS and LC-ESI-MS.
Central European Institute of Technology Masaryk University 625 00 Brno Czech Republic
Department of Biosystems Science and Engineering ETH Zurich 4058 Basel Switzerland
Department of Chemistry and Applied Biosciences ETH Zurich 8093 Zurich Switzerland
Department of Chemistry Masaryk University 625 00 Brno Czech Republic
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