Purified angiotensin converting enzyme from Rana esculenta ovary influences ovarian steroidogenesis in vitro.
The aim of the present study was to purify and characterize angiotensin-converting enzyme (ACE) present in frog ovary (Rana esculenta). Detergent and trypsin-extracted enzymes were purified using a one-step process, consisting of affinity chromatography on lisinopril coupled to Sepharose 6B. The molecular mass was 150 kDa for both detergent-extracted and trypsin-extracted enzyme. The specific activity of detergent-extracted and trypsin-extracted ACE was 294 U mg(-1) and 326 U mg(-1) respectively. The optimum pH range was from 7-8.5 at 37 degrees C and the optimum temperature was 50 degrees C. Optimum chloride concentration was about 200 mM for synthetic substrate FAPGG (N-[3-(2-furyl)acryloyl] L-phenylalanyl glycyl glycine) and angiotensin I, and 10 mM for bradykinin. The Km and Kcat values for FAPGG were 0.608 +/- 0.07 mM and 249 sec(-1) respectively and I50 values for captopril and lisinopril, two specific ACE inhibitors, were 68 +/- 12.55 nM and 6.763 +/- 0.66 nM respectively. Frog ovary tissue from prereproductive period was incubated in vitro in the presence of frog ovary ACE (2.5 mU/ml), captopril (0.1 mM), and lisinopril (0.1 mM). Production of 17beta-estradiol, progesterone, and prostaglandins E2 and F2alpha was determined. The data showed a modulation of 17beta-estradiol, progesterone and prostaglandin E2 production by ovary ACE.
Miano
A
A
Department of Molecular, Cellular and Animal Biology, University of Camerino, I-62032 Camerino, Italy.
Quassinti
L
L
Maccari
E
E
Murri
O
O
Amici
D
D
Bramucci
M
M
D000803:Angiotensin I / Q000378:metabolism; D000806:Angiotensin-Converting Enzyme Inhibitors / Q000378:metabolism; D000818:Animals; D001920:Bradykinin / Q000378:metabolism; D002216:Captopril / Q000378:metabolism; D005260:Female; D006863:Hydrogen-Ion Concentration; D017706:Lisinopril / Q000378:metabolism; D009842:Oligopeptides / Q000378:metabolism; D010053:Ovary / Q000201:enzymology*; D011893:Rana esculenta / Q000033:anatomy & histology / Q000378:metabolism*; D012083:Renin / Q000302:isolation & purification* / Q000378:metabolism; D012965:Sodium Chloride / Q000378:metabolism; D013256:Steroids / Q000096:biosynthesis*; D013696:Temperature
D000806:Angiotensin-Converting Enzyme Inhibitors; D009842:Oligopeptides; D013256:Steroids; D012965:Sodium Chloride; C038275:2-furanacryloyl-phenylalanyl-glycyl-glycine; D000803:Angiotensin I; D002216:Captopril; D017706:Lisinopril; D012083:Renin; D001920:Bradykinin