UNIT 17.19B Composition of Labeled Monosaccharides from Glycosaminoglycans
Published Online: 1 MAY 2001
Copyright © 2003 by John Wiley and Sons, Inc.
Lab Protocol Title
Current Protocols in Molecular Biology
How to Cite
Conrad, H. E. 2001. Composition of Labeled Monosaccharides from Glycosaminoglycans. Current Protocols in Molecular Biology. 32:IV:17.19B:17.19.26–17.19.32.
- Published Online: 1 MAY 2001
- Published Print: OCT 1995
Proteoglycans can be metabolically labeled with [3H]glucosamine to incorporate label into glucosamine and galactosamine. Glycosaminoglycans can be released from the proteoglycans and the distribution of labeled glucosamine and galactosamine in the isolated glycosaminoglycans can be determined. A series of acid treatments yields a mixture of disaccharides (uronosyl-anhydro sugars), free uronic acids, and free anhydro sugars, which are then reduced and separated by paper chromatography. The amount of each labeled component is quantified by scintillation counting. This procedure can also be used to determine the composition of glycosaminoglycans that have not been labeled metabolically. Samples of unlabeled glycosaminoglycans can be treated in the same manner and the hydrolysis/deamination mixture can be reduced with NaB3H4 to give stoichiometric 3H labeling of each reducing sugar component of the mixture.