Purification of normal human urinary N-acetyl-beta-hexosaminidase A by affinity chromatography.
N-Acetyl-beta-hexosaminidase A was purified 1000-fold from human urine by chromatography on DEAE-Sephadex followed by concanavalin A--Sepharose affinity chromatography. The optimal pH range was 4.4--4.5 for both the N-acetylglucosamine and N-acetylgalactosamine derivatives. The Km values were 0.51 mM and 0.28 mM respectively for the N-acetylglucosamine and N-acetylgalactosamine derivatives. The glycoprotein nature of the urinary enzyme was established by its affinity towards concanavalin A as well as by the presence of sialic acid, galactose, glucose, mannose and hexosamines in the molecule.
D002241:Carbohydrates / Q000032:analysis; D002846:Chromatography, Affinity; D002848:Chromatography, DEAE-Cellulose; D003208:Concanavalin A; D004591:Electrophoresis, Polyacrylamide Gel; D006596:Hexosaminidases / Q000032:analysis / Q000037:antagonists & inhibitors / Q000302:isolation & purification* / Q000652:urine; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D013696:Temperature
D002241:Carbohydrates; D003208:Concanavalin A; D006596:Hexosaminidases