Differential Isotope Labeling of Glycopeptides for Accurate Determination of Differences in Site-Specific Glycosylation

• Autoři: PABST Martin; BENEŠOVÁ Iva; FAGERER Stephan R; JACOBSEN Mathias; EYER Klaus; SCHMIDT Gregor; STEINHOFF Robert; KRISMER Jasmin; WAHL Fabian; PREISLER Jan; ZENOBI Renato
• Rok publikování: 2016
• Druh: Článek v odborném periodiku
• Časopis / Zdroj: Journal of Proteome Research
• Fakulta / Pracoviště MU: Přírodovědecká fakulta
• Doi: http://dx.doi.org/10.1021/acs.jproteome.5b00899
• Obor: Analytická chemie, separace
• Klíčová slova: glycopeptides; stable isotope labeling; IgG; MALDI MS; LC-ESI-MS
• Popis: 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 ((D4C4)-C-13) provides an 8 Da mass increment over the light natural form ((H4C4)-C-12), 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 IgGI Pc glycopeptides from polyclonal IgG samples with respect to their galactosylation and sialylation patterns using MALDI MS and LC-ESI-MS.

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• CEITEC - středoevropský technologický institut
• Zaměstnáním čerstvých absolventů doktorského studia k vědecké excelenci