Isolation and Characterization of the Rhesus Monkey Renal 15-Hydroxy Prostaglandin Dehydrogenase
Lambert, Jack S.
MetadataShow full item record
Isolation of the Rhesus monkey renal 15-hydroxydehydrogenase resulted in an approximately 85-fold purification by using DEAE-cellulose and hydroxylapatite chromatography purification methods. Disc gel electrophoresis showed several protein bands, indicating the partial purity of the enzyme. Assay studies indicated that the specific cofactor for the enzyme was NAD+; NADP+ was not a cofactor. The best substrates were PGE1, PGA1, PGE2 and PGA2, as indicated by Km of 8.0 ~M, 7.5 ~M, 7.8 ~M, and 9.26 ~M, respectively. The prostaglandin analogs 17-~-PGE1 and ~4-PGE1 had Km of 12.2 ~M and 27.4 ~M, respectively; with the PGF1a and PGF2a being substrates to a lesser degree, 31.25 ~M and 62.5 ~M. The following compounds were poor substrates or not substrates at all: PGB1, PGB2, PGD1' 5-oxa PGF1a, l5(R) PGE2' 16,16-dimethyl PGFla, and 15-methy1 PGE2. Of the inhibitors tested, ethacrynic acid, furosamide, and prostanoic acid exhibited inhibition, whereas angiotensin II, eledoisin, and bradykinin did not. The results suggest a successful method for enzyme purification, show the specificity of the enzyme, and help elucidate the mode of action of certain known inhibitors of renal function.