Purification and immunochemical characterization of the cytoplasmic androgen-binding protein of rat liver.
The cytoplasmic androgen-binding (CAB) protein of the male rat liver has been implicated to play a role in the androgen-dependent regulation of alpha 2u-globulin synthesis. The liver of the adult male rat contains about 50 fmol of specific high-affinity androgen-binding activity per milligram of total cytosolic protein. Photoaffinity labeling with [3H]R-1881 followed by SDS-polyacrylamide gel electrophoresis and autoradiography shows that the CAB is a 31-kilodalton protein. By means of DEAE-cellulose chromatography and preparative SDS-polyacrylamide gel electrophoresis, we have purified the CAB protein to electrophoretic homogeneity and have raised polyclonal rabbit antiserum that is monospecific to this protein. In the sucrose density gradient, the antiserum reacted with the androgen-binding component of the male liver cytosol prelabeled with tritiated dihydrotestosterone. Western blot analysis of the liver cytosol showed that the antiserum recognizes only the 31-kDa androgen-binding component. Such immunoblotting also showed that unlike the young adult, the androgen-insensitive states during prepuberty and senescence are associated with a marked reduction in the hepatic concentration of the immunoreactive CAB protein. No immuno-chemical cross-reactivity between CAB and another androgen-binding component of Mr 29K (which is associated with androgen insensitivity during prepuberty and senescence) was observed. The latter finding favors the possibility that 31- and 29-kDa androgen-binding components may have distinct sequence structure.
Demyan
W F
WF
Department of Obstetrics and Gynecology, University of Texas Health Science Center, San Antonio 78284.
Sarkar
F H
FH
Murty
C V
CV
Roy
A K
AK
D000375:Aging; D000727:Androgen-Binding Protein / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism; D000818:Animals; D000906:Antibodies; D000937:Antigen-Antibody Reactions; D002499:Centrifugation, Density Gradient; D003600:Cytosol / Q000378:metabolism; D004963:Estrenes / Q000378:metabolism; D005260:Female; D008099:Liver / Q000254:growth & development / Q000378:metabolism*; D008297:Male; D015741:Metribolone; D008970:Molecular Weight; D051381:Rats; D012737:Sex Factors
D000727:Androgen-Binding Protein; D000906:Antibodies; D004963:Estrenes; D015741:Metribolone