title: Characterization of a hepatic protein in nonhuman primates that binds mibolerone but not dihydrotestosterone or methyltrienolone.
abstract: During our studies of the hepatic androgen receptor in cynomolgus monkeys, tritiated mibolerone +/- a 200-fold excess of unlabeled mibolerone has been used to determine specific binding in cytosol. During time-course studies, high-capacity, unsaturable binding of [3H]mibolerone was noted after short-term incubations (4 h, 4 degrees C). When hepatic cytosol from male monkeys was incubated for 18 h at 4 degrees C, the high-capacity binding disappeared; saturable, high-affinity binding with characteristics consistent with the androgen receptor then could be identified. The characterization of [3H]mibolerone binding in molybdate-stabilized hepatic cytosol using sucrose density gradients and gel filtration yielded an unstable binding peak in addition to that of the androgen receptor. This lower molecular weight protein identified by gel filtration did not bind other androgens, including methyltrienolone, and did not have characteristics of other binding proteins that have been identified previously. This protein was not precipitated from 30% ammonium sulfate, which allowed it to be separated from the androgen receptor. Binding to this protein in ovariectomized female monkeys did not disappear with extended incubation at 4 degrees C, suggesting greater stability or a higher capacity. The function of this protein is not known, but both triamcinolone acetonide and contraceptive progestins appeared to displace tritiated mibolerone that was bound to it. This high-capacity binding of mibolerone interferes in the assessment of androgen receptor levels in these females unless it is eliminated. The synthetic androgen methyltrienolone does not bind to this protein and is a better choice for defining binding to the androgen receptor in these tissues.
authors.affiliation: Fort Wayne Center for Medical Education, Indiana University Medical School 46805, USA.
authors.forename: D R
authors.initials: DR
authors.lastname: Koritnik
authors.forename: K B
authors.initials: KB
authors.lastname: Marschke
authors.forename: A
authors.initials: A
authors.lastname: Koshy
mesh_terms: D000645:Ammonium Sulfate / Q000737:chemistry; D000818:Animals; D001667:Binding, Competitive; D011232:Chemical Precipitation; D002846:Chromatography, Affinity; D002853:Chromatography, Liquid; D003600:Cytosol / Q000737:chemistry / Q000378:metabolism; D013196:Dihydrotestosterone / Q000737:chemistry / Q000378:metabolism; D004958:Estradiol / Q000378:metabolism / Q000494:pharmacology; D005260:Female; D008099:Liver / Q000737:chemistry*; D008252:Macaca fascicularis; D008297:Male; D015741:Metribolone / Q000737:chemistry / Q000378:metabolism; D015394:Molecular Structure; D009277:Nandrolone / Q000031:analogs & derivatives* / Q000378:metabolism; D010052:Ovariectomy; D011506:Proteins / Q000032:analysis / Q000737:chemistry / Q000378:metabolism*; D011944:Receptors, Androgen / Q000737:chemistry / Q000378:metabolism*
chemical_list: D011506:Proteins; D011944:Receptors, Androgen; D013196:Dihydrotestosterone; D015741:Metribolone; D004958:Estradiol; D009277:Nandrolone; C100075:mibolerone; D000645:Ammonium Sulfate