Purification and characterization of rat brain prostaglandin D synthetase.

摘要: Prostaglandin D synthetase was purified 2,600-fold from rat brain to apparent homogeneity, as judged by polyacrylamide gel electrophoresis and ultracentrifugation. The enzyme a monomeric protein with molecular weight of 27,000 +/- 1,000. pI value, sedimentation coefficient, partial specific volume were 4.6, 4.1 s, 0.73 ml/g, respectively. stable between pH 4 11 at the temperature lower than 25 degrees C resistant heat treatment under alkaline conditions (pH 8-11). About 50% activity detected after 100 for 5 min 10. However, readily inactivated isomerase reaction prostaglandin H2 D2. required sulfhydryl compounds such dithiothreitol, glutathione, beta-mercaptoethanol, cysteine, cysteamine reaction, but stoichiometric oxidation these not observed. optimum pH, Km value H2, turnover number 9.5, 14 microM, 170 min-1, antibody raised against in rabbit, which showed only one positive band immunoblotting crude extracts same position that enzyme. More 90% absorbed an excess amount antibody, indicating our preparation is major component responsible biosynthesis D2 brain.