PABST, Martin, Iva BENEŠOVÁ, Stephan R FAGERER, Mathias JACOBSEN, Klaus EYER, Gregor SCHMIDT, Robert STEINHOFF, Jasmin KRISMER, Fabian WAHL, Jan PREISLER and Renato ZENOBI. Differential Isotope Labeling of Glycopeptides for Accurate Determination of Differences in Site-Specific Glycosylation. Journal of Proteome Research. Washington: American Chemical Society, 2016, vol. 15, No 1, p. 326-331. ISSN 1535-3893. Available from: https://dx.doi.org/10.1021/acs.jproteome.5b00899.
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Basic information
Original name Differential Isotope Labeling of Glycopeptides for Accurate Determination of Differences in Site-Specific Glycosylation
Authors PABST, Martin (40 Austria), Iva BENEŠOVÁ (203 Czech Republic, belonging to the institution), Stephan R FAGERER (276 Germany), Mathias JACOBSEN (756 Switzerland), Klaus EYER (756 Switzerland), Gregor SCHMIDT (756 Switzerland), Robert STEINHOFF (276 Germany), Jasmin KRISMER (756 Switzerland), Fabian WAHL (756 Switzerland), Jan PREISLER (203 Czech Republic, guarantor, belonging to the institution) and Renato ZENOBI (756 Switzerland).
Edition Journal of Proteome Research, Washington, American Chemical Society, 2016, 1535-3893.
Other information
Original language English
Type of outcome Article in a journal
Field of Study 10406 Analytical chemistry
Country of publisher United States of America
Confidentiality degree is not subject to a state or trade secret
Impact factor Impact factor: 4.268
RIV identification code RIV/00216224:14310/16:00093930
Organization unit Faculty of Science
Doi http://dx.doi.org/10.1021/acs.jproteome.5b00899
UT WoS 000367706000026
Keywords in English glycopeptides; stable isotope labeling; IgG; MALDI MS; LC-ESI-MS
Tags AKR, rivok
Tags International impact, Reviewed
Changed by Changed by: prof. Mgr. Jan Preisler, Ph.D., učo 45329. Changed: 15/2/2018 22:03.
Abstract
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.
Links
ED1.1.00/02.0068, research and development projectName: CEITEC - central european institute of technology
EE2.3.30.0009, research and development projectName: Zaměstnáním čerstvých absolventů doktorského studia k vědecké excelenci
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