A Visualizable Chain-Terminating Inhibitor of Glycosaminoglycan Biosynthesis in Developing Zebrafish

Heparan sulfate (HS) and chondroitin sulfate (CS) glycosaminoglycans (GAG) are proteoglycan‐associated polysaccharides with essential functions in animals. They have been studied extensively by genetic manipulation of biosynthetic enzymes, but chemical tools for probing GAG function are limited. HS...

Full description

Saved in:
Bibliographic Details
Published in:Angewandte Chemie International Edition 2014-03, Vol.53 (13), p.3347-3352
Main Authors: Beahm, Brendan J., Dehnert, Karen W., Derr, Nicolas L., Kuhn, Joachim, Eberhart, Johann K., Spillmann, Dorothe, Amacher, Sharon L., Bertozzi, Carolyn R.
Format: Article
Language:English
Subjects:
Animals
Biosynthesis
Chains
Chondroitin Sulfates - chemistry
Click Chemistry
Glycosaminoglycans
Glycosaminoglycans - metabolism
Heparan sulfate
Heparitin Sulfate - chemistry
Inhibitors
Residues
Sulfates
Xylose
Zebrafish
Zebrafish - metabolism
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Heparan sulfate (HS) and chondroitin sulfate (CS) glycosaminoglycans (GAG) are proteoglycan‐associated polysaccharides with essential functions in animals. They have been studied extensively by genetic manipulation of biosynthetic enzymes, but chemical tools for probing GAG function are limited. HS and CS possess a conserved xylose residue that links the polysaccharide chain to a protein backbone. Here we report that, in zebrafish embryos, the peptide‐proximal xylose residue can be metabolically replaced with a chain‐terminating 4‐azido‐4‐deoxyxylose (4‐XylAz) residue by administration of UDP‐4‐azido‐4‐deoxyxylose (UDP‐4‐XylAz). UDP‐4‐XylAz disrupted both HS and CS biosynthesis and caused developmental abnormalities reminiscent of GAG biosynthesis and laminin mutants. The azide substituent of protein‐bound 4‐XylAz allowed for rapid visualization of the organismal sites of chain termination in vivo through bioorthogonal reaction with fluorescent cyclooctyne probes. UDP‐4‐XylAz therefore complements genetic tools for studies of GAG function in zebrafish embryogenesis. Visualizing inhibition: Metabolic incorporation of an azide modified xylose residue inhibits elaboration of the glycan. Further, the azide group enables rapid visualization of the sites of inhibition in vivo during zebrafish development using Cu‐free click chemistry.
ISSN:1433-7851
1521-3773
1521-3773
DOI:10.1002/anie.201310569