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

J 2016

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

PABST, Martin, Iva BENEŠOVÁ, Stephan R FAGERER, Mathias JACOBSEN, Klaus EYER et. al.

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

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

10406 Analytical chemistry

Country of publisher

United States of America

Confidentiality degree

není předmětem státního či obchodního tajemství

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

Abstract

V originále

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 project

Name: CEITEC - central european institute of technology

EE2.3.30.0009, research and development project

Name: Zaměstnáním čerstvých absolventů doktorského studia k vědecké excelenci

Citovat

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.

@article{1375802,
   author = {Pabst, Martin and Benešová, Iva and Fagerer, Stephan R and Jacobsen, Mathias and Eyer, Klaus and Schmidt, Gregor and Steinhoff, Robert and Krismer, Jasmin and Wahl, Fabian and Preisler, Jan and Zenobi, Renato},
   article_location = {Washington},
   article_number = {1},
   doi = {http://dx.doi.org/10.1021/acs.jproteome.5b00899},
   keywords = {glycopeptides; stable isotope labeling; IgG; MALDI MS; LC-ESI-MS},
   language = {eng},
   issn = {1535-3893},
   journal = {Journal of Proteome Research},
   title = {Differential Isotope Labeling of Glycopeptides for Accurate Determination of Differences in Site-Specific Glycosylation},
   volume = {15},
   year = {2016}
}

TY  - JOUR
ID  - 1375802
AU  - Pabst, Martin - Benešová, Iva - Fagerer, Stephan R - Jacobsen, Mathias - Eyer, Klaus - Schmidt, Gregor - Steinhoff, Robert - Krismer, Jasmin - Wahl, Fabian - Preisler, Jan - Zenobi, Renato
PY  - 2016
TI  - Differential Isotope Labeling of Glycopeptides for Accurate Determination of Differences in Site-Specific Glycosylation
JF  - Journal of Proteome Research
VL  - 15
IS  - 1
SP  - 326-331
EP  - 326-331
PB  - American Chemical Society
SN  - 15353893
KW  - glycopeptides
KW  - stable isotope labeling
KW  - IgG
KW  - MALDI MS
KW  - LC-ESI-MS
N2  - 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.
ER  -

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. \textit{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.

Detailed Information on Publication Record