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

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.

authors

Dube DH,Prescher JA,Quang CN,Bertozzi CR

doi

10.1073/pnas.0506855103

keywords:

subject

Has Abstract

pub_date

2006-03-28 00:00:00

pages

4819-24

issue

13

eissn

0027-8424

issn

1091-6490

pii

0506855103

journal_volume

103

pub_type

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