Activation of human liver 3 alpha-hydroxysteroid dehydrogenase by sulphobromophthalein.
Human liver contains at least two isoenzymes (DD2 and DD4) of 3 alpha-hydroxysteroid/dihydrodiol dehydrogenase. The NADP(H)-linked oxidoreductase activities of DD4 were activated more than 4-fold by sulphobromophthalein at concentrations above 20 microM and under physiological pH conditions. Sulphobromophthalein did not stimulate the activities of DD2 and human liver aldehyde reductase, which are functionally and/or structurally related to DD4. No stimulatory effect on the activity of DD4 was observed with other organic anions such as Indocyanine Green, haematin and Rose Bengal. The binding of sulphobromophthalein to DD4 was instantaneous and reversible, and was detected by fluorescence and ultrafiltration assays. The activation by sulphobromophthalein decreased the activation energy in the dehydrogenation reaction for the enzyme, and increased both kcat, and Km values for the coenzymes and substrates. Kinetic analyses with respect to concentrations of NADP+ and (S)-(+)-indan-1-ol indicated that sulphobromophthalein was a non-essential activator of mixed type showing a dissociation constant of 2.6 microM. Thus, the human 3 alpha-hydroxysteroid dehydrogenase isoenzyme has a binding site specific to sulphobromophthalein, and the hepatic metabolism mediated by this isoenzyme may be influenced when this drug is administered.
Biochemistry Laboratory, Gifu Pharmaceutical Univeristy, Japan.
K
K
Matsuura
Y
Y
Tamada
Y
Y
Deyashiki
Y
Y
Miyabe
M
M
Nakanishi
I
I
Ohya
A
A
Hara
D015096:3-Hydroxysteroid Dehydrogenases / Q000187:drug effects* / Q000378:metabolism*; D045002:3-alpha-Hydroxysteroid Dehydrogenase (B-Specific); D000429:Alcohol Oxidoreductases / Q000378:metabolism; D001665:Binding Sites; D004789:Enzyme Activation; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008099:Liver / Q000187:drug effects* / Q000201:enzymology*; D010088:Oxidoreductases; D044925:Oxidoreductases Acting on CH-CH Group Donors; D012680:Sensitivity and Specificity; D013448:Sulfobromophthalein / Q000378:metabolism / Q000494:pharmacology*
D013448:Sulfobromophthalein; D010088:Oxidoreductases; D015096:3-Hydroxysteroid Dehydrogenases; D000429:Alcohol Oxidoreductases; C023125:dihydrodiol dehydrogenases; D045002:3-alpha-Hydroxysteroid Dehydrogenase (B-Specific); D044925:Oxidoreductases Acting on CH-CH Group Donors; C116788:cis-1,2-dihydro-1,2-dihydroxynaphthalene dehydrogenase