Prostaglandin-E2 9-ketoreductase from swine kidney. Production of antisera and application to development of a radioimmunoassay.
Prostaglandin-E2 9-ketoreductase (PGE2-9-KR, EC 1.1.1.189), the enzyme which catalyzes the reaction from prostaglandin E2 (PGE2) to prostaglandin F2 alpha (PGF2 alpha), was purified 580-fold from swine kidney. The molecular mass of the enzyme determined by SDS-gel electrophoresis was 33 kDa. Antiserum against the purified enzyme was raised in three rabbits. The antiserum was able to precipitate PGE2-9-KR from swine kidney and to crossreact with pGE2-9-KR from several reproductive organ tissues, such as rabbit ovary, rabbit corpus luteum, rabbit endometrium and human decidua vera. When swine kidney PGE2-9-KR was labelled with 125I and incubated with affinity-purified antiserum in the presence of increasing amounts of unlabelled enzyme, competitive binding of the unlabelled enzyme to the antibody was observed. A radioimmunoassay for the quantitation of the enzyme was developed. The standard curve was linear from 5 to 500 ng enzyme. The intra- and interassay coefficients of variation were 6.4 and 13.2%, respectively. The assay may be useful for the quantitation of PGE2-9-KR in several tissues under various physiological conditions.
Krüger
S
S
Universitäts-Frauenklinik, Münster, F.R.G.
Meyer
U
U
Daniels
D
D
Schlegel
W
W
D000818:Animals; D003429:Cross Reactions; D003656:Decidua / Q000201:enzymology; D005260:Female; D006911:Hydroxyprostaglandin Dehydrogenases / Q000302:isolation & purification*; D007106:Immune Sera; D007668:Kidney / Q000201:enzymology*; D010053:Ovary / Q000201:enzymology; D011863:Radioimmunoassay / Q000379:methods; D013552:Swine
D007106:Immune Sera; D006911:Hydroxyprostaglandin Dehydrogenases; C035858:prostaglandin-E2 9-reductase