Hydrophobic-interaction chromatography of glycosaminoglycuronans: the contribution of N-acetyl groups in heparin and heparan sulfate to the affinity for hydrophobic gels, and variety of molecular species in beef-kidney heparan sulfate.
Contribution of N-acetyl groups in heparin and heparan sulfate to their affinity for hydrophobic gels was examined by use of a series of semi-synthetic, N-acetylated, hog-intestinal heparins, a whale-intestinal heparin, and a beef-kidney heparan sulfate. Chromatography on Phenyl-Sepharose CL-4B in 3.8-1.0M ammonium sulfate-10mM hydrochloric acid indicated that an increasing N-acetyl content, which is correlated to a decreasing N-sulfate content, results in a marked increase in the affinity for the gels. The variety of molecular species in beef-kidney heparan sulfate, previously fractionated by conventional chromatographic procedures, was demonstrated by separating further, by hydrophobic-interaction chromatography, the polysaccharide into several fractions composed of molecular species distinctly different in N-acetyl and sulfate content, and in molecular size.
D000818:Animals; D002417:Cattle; D002846:Chromatography, Affinity / Q000379:methods; D002850:Chromatography, Gel / Q000379:methods; D006025:Glycosaminoglycans / Q000302:isolation & purification*; D006493:Heparin / Q000302:isolation & purification*; D006497:Heparan Sulfate / Q000302:isolation & purification*; D007422:Intestines / Q000032:analysis; D007668:Kidney / Q000032:analysis*; D012685:Sepharose / Q000031:analogs & derivatives; D013329:Structure-Activity Relationship; D014907:Whales
D006025:Glycosaminoglycans; D006493:Heparin; D006497:Heparan Sulfate; D012685:Sepharose; C018303:glycosaminoglucuronates; C033452:Phenyl-Sepharose CL-4B