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Title: Characterization of protein N-glycosylation by tandem mass spectrometry using complementary fragmentation techniques

Abstract

The analysis of post-translational modifications (PTMs) by proteomics is regarded as a technically challenging undertaking. While in recent years approaches to examine and quantify protein phosphorylation have greatly improved, the analysis of many protein modifications, such as glycosylation, are still regarded as problematic. Limitations in the standard proteomics workflow, such as use of suboptimal peptide fragmentation methods, can significantly prevent the identification of glycopeptides. The current generation of tandem mass spectrometers has made available a variety of fragmentation options, many of which are becoming standard features on these instruments. Lastly, we have used three common fragmentation techniques, namely CID, HCD, and ETD, to analyze a glycopeptide and highlight how an integrated fragmentation approach can be used to identify the modified residue and characterize the N-glycan on a peptide.

Authors:
 [1];  [1];  [2];  [1]
  1. Univ. of Melbourne, Melbourne, VIC (Australia)
  2. Univ. of Melbourne, Melbourne, VIC (Australia); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Publication Date:
Research Org.:
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23)
OSTI Identifier:
1257263
Grant/Contract Number:  
CE110001007; FT130101165; AC02-05CH11231
Resource Type:
Accepted Manuscript
Journal Name:
Frontiers in Plant Science
Additional Journal Information:
Journal Volume: 6; Journal ID: ISSN 1664-462X
Publisher:
Frontiers Research Foundation
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; glycosylation; ragmentation; electron-transfer dissociation; post-translational modification; tandem mass spectrometry

Citation Formats

Ford, Kristina L., Zeng, Wei, Heazlewood, Joshua L., and Bacic, Antony. Characterization of protein N-glycosylation by tandem mass spectrometry using complementary fragmentation techniques. United States: N. p., 2015. Web. doi:10.3389/fpls.2015.00674.
Ford, Kristina L., Zeng, Wei, Heazlewood, Joshua L., & Bacic, Antony. Characterization of protein N-glycosylation by tandem mass spectrometry using complementary fragmentation techniques. United States. doi:10.3389/fpls.2015.00674.
Ford, Kristina L., Zeng, Wei, Heazlewood, Joshua L., and Bacic, Antony. Fri . "Characterization of protein N-glycosylation by tandem mass spectrometry using complementary fragmentation techniques". United States. doi:10.3389/fpls.2015.00674. https://www.osti.gov/servlets/purl/1257263.
@article{osti_1257263,
title = {Characterization of protein N-glycosylation by tandem mass spectrometry using complementary fragmentation techniques},
author = {Ford, Kristina L. and Zeng, Wei and Heazlewood, Joshua L. and Bacic, Antony},
abstractNote = {The analysis of post-translational modifications (PTMs) by proteomics is regarded as a technically challenging undertaking. While in recent years approaches to examine and quantify protein phosphorylation have greatly improved, the analysis of many protein modifications, such as glycosylation, are still regarded as problematic. Limitations in the standard proteomics workflow, such as use of suboptimal peptide fragmentation methods, can significantly prevent the identification of glycopeptides. The current generation of tandem mass spectrometers has made available a variety of fragmentation options, many of which are becoming standard features on these instruments. Lastly, we have used three common fragmentation techniques, namely CID, HCD, and ETD, to analyze a glycopeptide and highlight how an integrated fragmentation approach can be used to identify the modified residue and characterize the N-glycan on a peptide.},
doi = {10.3389/fpls.2015.00674},
journal = {Frontiers in Plant Science},
number = ,
volume = 6,
place = {United States},
year = {2015},
month = {8}
}

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Cited by: 8 works
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