We thank J. Jewett, E. Sletten, B. Belardi, J. Hudak, T. Gallagher, S. Laughlin and B. Swarts for helpful discussions and for critical reading of the manuscript. This work was supported by NIH grants to C.R.B. (GM58867) and S.L.A. (GM61952). The lamb1ab1166 allele was isolated in a screen supported by NIH grant HD22486 (to Charles B. Kimmel).
A Visualizable Chain-Terminating Inhibitor of Glycosaminoglycan Biosynthesis in Developing Zebrafish†
Article first published online: 19 FEB 2014
© 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Angewandte Chemie International Edition
Volume 53, Issue 13, pages 3347–3352, March 24, 2014
How to Cite
Beahm, B. J., Dehnert, K. W., Derr, N. L., Kuhn, J., Eberhart, J. K., Spillmann, D., Amacher, S. L. and Bertozzi, C. R. (2014), A Visualizable Chain-Terminating Inhibitor of Glycosaminoglycan Biosynthesis in Developing Zebrafish. Angew. Chem. Int. Ed., 53: 3347–3352. doi: 10.1002/anie.201310569
- Issue published online: 19 MAR 2014
- Article first published online: 19 FEB 2014
- Manuscript Received: 5 DEC 2013
- click chemistry;
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.