Evolutionary differences in glycosaminoglycan fine structure detected by quantitative glycan reductive isotope labeling.
To facilitate qualitative and quantitative analysis of glycosaminoglycans, we tagged the reducing end of lyase-generated disaccharides with aniline-containing stable isotopes (12C6 and 13C6). Because different isotope tags have no effect on chromatographic retention times but can be discriminated by a mass detector, differentially isotope-tagged samples can be compared simultaneously by liquid chromatography/mass spectrometry and quantified by admixture with known amounts of standards. The technique is adaptable to all types of glycosaminoglycans, and its sensitivity is only limited by the type of mass spectrometer available. We validated the method using commercial heparin and keratan sulfate as well as heparan sulfate isolated from mutant and wild-type Chinese hamster ovary cells, and select tissues from mutant and wild-type mice. This new method provides more robust, reliable, and sensitive means of quantitative evaluation of glycosaminoglycan disaccharide compositions than existing techniques allowing us to compare the chondroitin and heparan sulfate compositions of Hydra vulgaris, Drosophila melanogaster, Caenorhabditis elegans, and mammalian cells. Our results demonstrate significant differences in glycosaminoglycan structure among these organisms that might represent evolutionarily distinct functional motifs.
Lawrence
Roger
R
Department of Cellular and Molecular Medicine, Glycobiology Research and Training Center, University of California, San Diego, La Jolla, California 92093, USA.
Olson
Sara K
SK
Steele
Robert E
RE
Wang
Lianchun
L
Warrior
Rahul
R
Cummings
Richard D
RD
Esko
Jeffrey D
JD
D000818:Animals; D016466:CHO Cells; D017173:Caenorhabditis elegans / Q000737:chemistry; D002236:Carbohydrate Conformation; D002247:Carbon Isotopes / Q000737:chemistry; D002853:Chromatography, Liquid / Q000379:methods; D006224:Cricetinae; D003412:Cricetulus; D004331:Drosophila melanogaster; D019143:Evolution, Molecular; D006025:Glycosaminoglycans / Q000737:chemistry*; D006829:Hydra / Q000737:chemistry; D007553:Isotope Labeling / Q000379:methods; D013058:Mass Spectrometry / Q000379:methods; D051379:Mice
D002247:Carbon Isotopes; D006025:Glycosaminoglycans