title: Production of size-defined heparosan, heparan sulfate, and heparin oligosaccharides by enzymatic depolymerization.
abstract: Glycosaminoglycans (GAG) are most commonly isolated as large polymers from various animal origins, the functional units of which are oligosaccharides, which bind their target proteins to induce conformational changes, compete with other ligands, or facilitate the formation of signaling complexes. One example, the extensively studied heparin pentasaccharide sequence-which binds antithrombin-III, inducing a conformational change that increases its serpin protease activity by 1,000-fold-is unique in that no other specific GAG-protein structure-function relations have been described to the same degree. Thus, production of heparan sulfate (HS) oligosaccharides is critical for obtaining specific structural information regarding the binding interactions of GAG and their ligands (typically proteins). Purely synthetic methods of oligosaccharide synthesis are possible, but the cost, time requirement, and difficulty of their preparation prohibit library synthesis in significant amounts. Herein, the use of bacterial heparin lyases for the production of HS oligosaccharides via enzymatic depolymerization of HS polymers is discussed. The separation and purification of these oligosaccharides by liquid chromatography are also described.
authors.affiliation: Department of Medicinal Chemistry, University of Utah, 30 South 2000 East, Skaggs #307, Salt Lake City, UT, 84112, USA.
authors.forename: Spencer
authors.initials: S
authors.lastname: Brown
authors.forename: Balagurunathan
authors.initials: B
authors.lastname: Kuberan
mesh_terms: D001671:Biochemistry / Q000379:methods*; D002498:Centrifugation; D002851:Chromatography, High Pressure Liquid; D004187:Disaccharides / Q000138:chemical synthesis* / Q000737:chemistry; D005374:Filtration; D006493:Heparin / Q000138:chemical synthesis* / Q000737:chemistry; D006497:Heparan Sulfate / Q000138:chemical synthesis* / Q000737:chemistry; D007474:Ion Exchange; D009844:Oligosaccharides / Q000138:chemical synthesis* / Q000737:chemistry; D058105:Polymerization; D011133:Polysaccharide-Lyases / Q000378:metabolism*; D013379:Substrate Specificity
chemical_list: D004187:Disaccharides; D006493:Heparin; D006497:Heparan Sulfate; D009844:Oligosaccharides; D011133:Polysaccharide-Lyases; C560363:heparosan; C021349:heparitinsulfate lyase