Purification and characterization of hypoxanthine/guanine phosphoribosyltransferase in bovine snout epidermis.
Hypoxanthine/guanine phosphoribosyltransferase was purified from bovine snout epidermis, about 600-fold by a combination method of centrifugation, ammonium sulfate fraction, Sephadex G-200 and DEAE cellulose chromatography. Enzymatic properties of the purified enzyme were determined as follows: pH optimum 7.2, temperature optimum 56 degrees C, and 82,000 in molecular weight. In the presence of phosphoribosyl pyrophosphate, the enzyme was extremely heat-stable. The enzyme displayed Michaelis-Menten kinetics with apparent Michaelis constants for hypoxanthine, guanine and phosphoribosyl pyrophosphate of 1.59, 20.4 and 72.6 microM respectively.
D000818:Animals; D002417:Cattle; D002498:Centrifugation; D005591:Chemical Fractionation; D002848:Chromatography, DEAE-Cellulose; D002852:Chromatography, Ion Exchange; D004817:Epidermis / Q000201:enzymology*; D006863:Hydrogen-Ion Concentration; D007041:Hypoxanthine Phosphoribosyltransferase / Q000032:analysis* / Q000302:isolation & purification; D007700:Kinetics; D008970:Molecular Weight; D013696:Temperature
D007041:Hypoxanthine Phosphoribosyltransferase