Purification and immunochemical characterization of the cytoplasmic androgen-binding protein of rat liver

Journal Article · Tue Feb 21 00:00:00 EST 1989 · Biochemistry; (USA)

DOI:https://doi.org/10.1021/bi00430a046· OSTI ID:5601316

Demyan, W F; Sarkar, F H; Ramana Murty, C V; Roy, A K ^[1]

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1. Univ. of Texas Health Science Center, San Antonio (USA)

The cytoplasmic androgen-binding (CAB) protein of the male rate liver has been implicated to play a role in the androgen-dependent regulation of {alpha}{sub 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 ({sup 3}H)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 SCS-polyacrylamide gel electrophoresis, the authors 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 immunochemical cross-reactivity between CAB and another androgen-binding component of M{sub r} 29K was observed. The latter finding favors the possibility that 31- and 29-kDa androgen-binding components may have distinct sequence structure.

• OSTI ID: 5601316
• Journal Information: Biochemistry; (USA), Vol. 28:4; ISSN 0006-2960
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
ANDROGENS
CROSS-LINKING
LIVER
CYTOCHEMISTRY
PROTEINS
AMINO ACID SEQUENCE
ELECTROPHORESIS
IMMUNE SERUMS
IMMUNOLOGY
PURIFICATION
RATS
TRITIUM COMPOUNDS
ANDROSTANES
ANIMALS
BIOCHEMISTRY
BODY
CHEMICAL REACTIONS
CHEMISTRY
DIGESTIVE SYSTEM
GLANDS
HORMONES
HYDROGEN COMPOUNDS
MAMMALS
MOLECULAR STRUCTURE
ORGANIC COMPOUNDS
ORGANS
POLYMERIZATION
RODENTS
STEROID HORMONES
STEROIDS
VERTEBRATES
550201* - Biochemistry- Tracer Techniques