Glutathione-prostaglandin A1 conjugate as substrate in the purification of prostaglandin 9-ketoreductase from rabbit kidney.
Using GSH-PGA1 as substrate for determination of enzyme activity a pI 4.8 form of rabbit kidney prostaglandin 9-keto-reductase has been purified 95 times to a specific activity of 1755 nmol/min per mg protein. The purification procedures involve ion-exchange chromatography, gel-filtration and affinity chromatography. The latter procedure comprises Blue Sepharose affinity chromatography, and GSH-PGA1-Sepharose affinity chromatography. The purified enzyme preparation also showed a weak NADP+-dependent 15-hydroxyprostaglandin dehydrogenase activity, 20 nmol/min per mg protein with PGE1 as substrate. Km (PGE1) for the dehydrogenase is 142.6 +/- 45.1 microM (S.E., n = 7).
D000818:Animals; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D005978:Glutathione / Q000378:metabolism* / Q000031:analogs & derivatives; D006911:Hydroxyprostaglandin Dehydrogenases / Q000302:isolation & purification* / Q000378:metabolism; D007526:Isoelectric Point; D007668:Kidney / Q000201:enzymology*; D008297:Male; D009249:NADP / Q000494:pharmacology; D011454:Prostaglandins A / Q000378:metabolism*; D011455:Prostaglandins A, Synthetic; D011817:Rabbits
D011454:Prostaglandins A; D011455:Prostaglandins A, Synthetic; D009249:NADP; C021031:GSH-prostaglandin A1; D006911:Hydroxyprostaglandin Dehydrogenases; D005978:Glutathione