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PMID: 16549800 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S.

Probing mucin-type O-linked glycosylation in living animals.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 103 ·No. 13 ·2006-03-28 ·Pages 4819-24

Dube DH, Prescher JA, Quang CN, Bertozzi CR

Abstract

Changes in O-linked protein glycosylation are known to correlate with disease states but are difficult to monitor in a physiological setting because of a lack of experimental tools. Here, we report a technique for rapid profiling of O-linked glycoproteins in living animals by metabolic labeling with N-azidoacetylgalactosamine (GalNAz) followed by Staudinger ligation with phosphine probes. After injection of mice with a peracetylated form of GalNAz, azide-labeled glycoproteins were observed in a variety of tissues, including liver, kidney, and heart, in serum, and on isolated splenocytes. B cell glycoproteins were robustly labeled with GalNAz but T cell glycoproteins were not, suggesting fundamental differences in glycosylation machinery or metabolism. Furthermore, GalNAz-labeled B cells could be selectively targeted with a phosphine probe by Staudinger ligation within the living animal. Metabolic labeling with GalNAz followed by Staudinger ligation provides a means for proteomic analysis of this posttranslational modification and for identifying O-linked glycoprotein fingerprints associated with disease.

MeSH Terms
Acetylgalactosamine/chemistry,pharmacology Animals Azides/chemistry,pharmacology Cell Line Glycosylation Hexosamines/chemistry,pharmacology Leukocytes/drug effects,metabolism Mice Molecular Structure Mucins/chemistry,metabolism
Chemicals
Azides Hexosamines Mucins N-azidoacetylmannosamine Acetylgalactosamine
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Dube Danielle H
Department of Chemistry, University of California, Berkeley, CA 94720, USA.
Prescher Jennifer A
Quang Chi N
Bertozzi Carolyn R
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.0506855103
Published
2006-03-28
Epub
2006-00-20
Pages
4819-24
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC1405625
Subset
IM
Grants
NIGMS NIH HHS · R01 GM066047 · United States
NIGMS NIH HHS · GM66047 · United States
External Links
PubMed source DOI: 10.1073/pnas.0506855103 PMC Full Text View journal details
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