Purification and characterization of mouse kidney beta-glucuronidase.

摘要: Beta-Glucuronidase has been purified from mouse kidneys previously induced by gonadotrophin to a specific enzyme activity 15 times higher than the non-induced kidney. The purification procedure includes ultrasonication solubilize enzyme, acid and ammonium sulfate precipitations, gel filtration in Sephadex G-200, DEAE-ion exchange chromatography, isoelectric focusing. resulting product of 284,000 Fishman units/mg protein, representing 1,090-fold is 17,000-fold level beta-glucuronidase apparently homogeneous criteria filtration, sodium dodecyl electrophoresis, immunodiffusion. Characterization showed that it identical with lysosomal isoenzymic electrophoretically, subunit molecular weight 74,000 (estimated electrophoresis) oligomer 300,000. stable at high temperature (up 55 degrees) wide range pH (from 4 11). It optimum for its 4.7 Km 1.18 10- minus M. characterization this kidney will have significance understanding nature isoenzymes be useful future studies on mechanism intracellular transport distribution hydrolase.