title: Apparent saturability of a 4S dihydrotestosterone-binding protein in rat muscle cytosol: role of 3 alpha-hydroxysteroid dehydrogenase and albumin.
abstract: In our studies of the binding of steroids in rat skeletal muscles, we have shown the existence of a saturable '4S' dihydrotestosterone-binding protein of low affinity (Kd = 1.16 X 10(-6) M). Competition studies led us to believe that the binding of dihydrotesterone (DHT) was due to 3 alpha-hydroxysteroid dehydrogenase (3 alpha-OHSD) enzyme responsible for the conversion of DHT to androstanediol (Adiol). Indeed, both the binding and the enzymatic activity are inhibited by various 3-keto steroids. In addition, the dissociation constant (Kd) and the Michaelis constant (Km) of the enzyme are similar. However, experiments with ammonium sulfate fractions of the cytosol have shown a partial separation of the binding and of the enzymatic activity. On the other hand, we have established that DHT (3 micrometer) is almost completely metabolized to Adiol during a 2-h incubation of 0 degrees C even in the absence of added coenzymes. Furthermore the '4S' protein binds Adiol more strongly than DHT and this binding is not saturable. Finally the binding behaviour of both DHT and Adiol with either muscle cytosol or rat albumin is similar when subjected to ammonium sulfate fractionation and sucrose density gradient centrifugation. In conclusion, the skeletal muscle 3 alpha-OHSD rapidly metabolizes DHT into Adiol which then binds strongly to a nonspecific binding protein, presumably rat serum albumin. Thus it can be said that the observed saturability of DHT binding is only apparent.
mesh_terms: D015096:3-Hydroxysteroid Dehydrogenases / Q000378:metabolism*; D015113:Androstane-3,17-diol / Q000378:metabolism; D000818:Animals; D003600:Cytosol / Q000378:metabolism; D013196:Dihydrotestosterone / Q000378:metabolism*; D005260:Female; D005333:Fetus / Q000378:metabolism; D007664:Ketosteroids / Q000494:pharmacology; D007700:Kinetics; D008297:Male; D009132:Muscles / Q000378:metabolism*; D011247:Pregnancy; D051381:Rats; D011944:Receptors, Androgen / Q000378:metabolism*; D011987:Receptors, Steroid / Q000378:metabolism*; D012709:Serum Albumin / Q000378:metabolism*; D013696:Temperature
chemical_list: D007664:Ketosteroids; D011944:Receptors, Androgen; D011987:Receptors, Steroid; D012709:Serum Albumin; D013196:Dihydrotestosterone; D015113:Androstane-3,17-diol; D015096:3-Hydroxysteroid Dehydrogenases