Different catalytic properties and inhibitor responses of the goldfish brain and ovary aromatase isozymes.
The brain and ovarian aromatase isozymes of goldfish (Carassius auratus) are encoded by different CYP19 genes. This study measured aromatase activity in the goldfish brain tissues. For a direct comparison of the properties of the two aromatase isozymes, Chinese hamster ovary cells were stably transfected with brain- and ovary-derived cDNAs (respectively, p450 arom B and -A) and the properties of the expressed isozymes were compared. The kinetic parameters of the two isozymes were determined using androstenedione and testosterone as substrates and compared to those of human aromatase. Inhibition profile analyses on the two isozymes were performed using seven inhibitors [4-hydroxyandrostenedione, 7 alpha-(4'-amino)phenylthio-1,4-androstadiene-3,17-dione, bridge (2,19-methyleneoxy)androstene-3,17-dione, aminoglutethimide (AG), CGS 20267, ICI D1033, and vorozole]. Except for AG, the compounds tested were found to be much stronger inhibitors against the ovary enzyme than the brain enzyme. In addition, the ovary isoform was more sensitive to two phytoestrogens, chrysin and 7,8-dihydroxyflavone, than the brain form. These studies reveal that catalytic properties of the goldfish aromatase isoforms are significantly different from those of human aromatase. In addition, differences in the K(i) values of aromatase inhibitors for the two goldfish isoforms suggest structural variance in the active sites of these isozymes.
Zhao
J
J
Division of Immunology, Beckman Research Institute of the City of the Hope, Duarte, California 91010, USA.
Mak
P
P
Tchoudakova
A
A
Callard
G
G
Chen
S
S
D000735:Androstenedione / Q000378:metabolism; D000818:Animals; D001141:Aromatase / Q000235:genetics / Q000378:metabolism*; D047072:Aromatase Inhibitors; D001921:Brain / Q000201:enzymology* / Q000648:ultrastructure; D016466:CHO Cells; D002384:Catalysis; D006224:Cricetinae; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D004968:Estrogens, Non-Steroidal / Q000494:pharmacology; D005260:Female; D005419:Flavonoids / Q000494:pharmacology; D015870:Gene Expression; D006054:Goldfish / Q000378:metabolism*; D007527:Isoenzymes / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D007529:Isoflavones; D007700:Kinetics; D008297:Male; D008861:Microsomes / Q000201:enzymology; D010053:Ovary / Q000201:enzymology*; D048789:Phytoestrogens; D028321:Plant Preparations; D013379:Substrate Specificity; D013739:Testosterone / Q000378:metabolism; D014162:Transfection
D047072:Aromatase Inhibitors; D004791:Enzyme Inhibitors; D004968:Estrogens, Non-Steroidal; D005419:Flavonoids; D007527:Isoenzymes; D007529:Isoflavones; D048789:Phytoestrogens; D028321:Plant Preparations; D013739:Testosterone; D000735:Androstenedione; D001141:Aromatase