Cellular O-Glycome Reporter/Amplification to explore O-glycans of living cells
File(s)
Author(s)
Type
Journal Article
Abstract
Protein O-glycosylation has key roles in many biological processes, but the repertoire of O-glycans synthesized by cells is difficult to determine. Here we describe an approach termed Cellular O-Glycome Reporter/Amplification (CORA), a sensitive method used to amplify and profile mucin-type O-glycans synthesized by living cells. Cells convert added peracetylated benzyl-α-N-acetylgalactosamine to a large variety of modified O-glycan derivatives that are secreted from cells, allowing for easy purification for analysis by HPLC and mass spectrometry (MS). Relative to conventional O-glycan analyses, CORA resulted in an ∼100-1,000-fold increase in sensitivity and identified a more complex repertoire of O-glycans in more than a dozen cell types from Homo sapiens and Mus musculus. Furthermore, when coupled with computational modeling, CORA can be used for predictions about the diversity of the human O-glycome and offers new opportunities to identify novel glycan biomarkers for human diseases.
Date Issued
2016-01-01
Date Acceptance
2015-10-26
Citation
Nature Methods, 2016, 13, pp.81-86
ISSN
1548-7105
Publisher
Nature Publishing Group
Start Page
81
End Page
86
Journal / Book Title
Nature Methods
Volume
13
Copyright Statement
© 2015 The Authors.
Sponsor
Wellcome Trust
Biotechnology and Biological Sciences Research Council (BBSRC)
Identifier
PII: nmeth.3675
Grant Number
102978/Z/13/Z
BB/K016164/1
Subjects
Science & Technology
Life Sciences & Biomedicine
Biochemical Research Methods
Biochemistry & Molecular Biology
SIALYL-LEWIS-X
PLURIPOTENT STEM-CELLS
MASS-SPECTROMETRY
IN-VITRO
CANCER
GLYCOSYLATION
TISSUES
DISEASE
ANTIGEN
COSMC
Animals
Chromatography, High Pressure Liquid
Mass Spectrometry
Mice
Polysaccharides
Animals
Mice
Polysaccharides
Chromatography, High Pressure Liquid
Mass Spectrometry
Developmental Biology
06 Biological Sciences
10 Technology
11 Medical and Health Sciences
Publication Status
Published
Date Publish Online
2015-11-30