title: Characterization of heparan sulfate from the unossified antler of Cervus elaphus.
abstract: The antler is the most rapidly growing tissue in the animal kingdom. According to previous reports, antler glycosaminoglycans (GAGs) consist of all kinds GAGs except for heparan sulfate (HS). Chondroitin sulfate is the major antler GAG component comprising 88% of the total uronic acid content. In the current study, we have isolated HS from antler for the first time and characterized it based on both NMR spectroscopy and disaccharide composition analysis. Antler GAGs were isolated by protease treatment and followed by cetylpyridinium chloride precipitation. The sensitivity of antler GAGs to heparin lyase III showed that this sample contained heparan sulfate. After incubation of antler GAGs with chondroitin lyase ABC, the HS-containing fraction was recovered by ethanol precipitation. The composition of HS disaccharides in this fraction was determined by its complete depolymerization with a mixture of heparin lyase I, II, and III and analysis of the resulting disaccharides by the reversed-phase (RP) ion pairing-HPLC, monitored by the fluorescence detection using 2-cyanoacetamide as a post-column labeling reagent. Eight unsaturated disaccharides (DeltaUA-GlcNAc, DeltaUA-GlcNS, DeltaUA-GlcNAc6S, DeltaUA2S-GlcNAc, DeltaUA-GlcNS6S, DeltaUA2S-GlcNS, DeltaUA2S-GlcNAc6S, DeltaUA2S-GlcNS6S) were produced from antler HS by digestion with the mixture of heparin lyases. The total content of 2-O-sulfo disaccharide units in antler HS was higher than that of heparan sulfate from most other animal sources.
authors.affiliation: Natural Products Research Institute, College of Pharmacy, Seoul National University, Seoul 110-460, Korea.
authors.forename: Young Wan
authors.initials: YW
authors.lastname: Ha
authors.forename: Byong Tae
authors.initials: BT
authors.lastname: Jeon
authors.forename: Sang Ho
authors.initials: SH
authors.lastname: Moon
authors.forename: Hidenao
authors.initials: H
authors.lastname: Toyoda
authors.forename: Toshihiko
authors.initials: T
authors.lastname: Toida
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Linhardt
authors.forename: Yeong Shik
authors.initials: YS
authors.lastname: Kim
mesh_terms: D000818:Animals; D000999:Antlers / Q000737:chemistry* / Q000378:metabolism; D003670:Deer / Q000378:metabolism*; D004187:Disaccharides / Q000032:analysis; D019763:Heparin Lyase / Q000737:chemistry; D006497:Heparan Sulfate / Q000737:chemistry* / Q000302:isolation & purification*; D009682:Magnetic Resonance Spectroscopy; D008297:Male
chemical_list: D004187:Disaccharides; D019763:Heparin Lyase; D006497:Heparan Sulfate