title: Androgen-binding protein. Purification from rat epididymis, characterization, and immunocytochemical localization.
abstract: Androgen-binding protein (ABP) was purified from caput epididymis of the rat by sequential chromatography on DEAE-Sepharose, hydroxylapatite, dihydrotestosterone-17 beta-hemisuccinyl-1,6-diaminohexane-Sepharose, and Sephadex G-150. The final product migrated as a single band corresponding to a peak of protein-bound [3H]dihydrotestosterone on polyacrylamide gel electrophoresis. A molecular weight of 100,000 was estimated by sedimentation equilibrium. On sodium dodecyl sulfate-polyacrylamide gel electrophoresis, subunits of Mr = 47,000 and 41,000 were observed. Amino acid analysis indicated ABP to be rich in leucine while nonpolar aminoacids totaled only 51%. Its carbohydrate content is 25%. Antibodies to purified ABP were raised in a rabbit and evaluated by immunodiffusion, immunoelectrophoresis, binding inhibition, radioimmunoassay, and immunocytochemistry. Immunoperoxidase staining localized ABP in the basal and adluminal regions of seminiferous tubules of rat testis and in secretory granules of cultured Sertoli cells. In principal cells of caput epididymis, ABP is concentrated in the supranuclear region known to contain morphological specializations for absorption. These immunocytochemical results confirm that ABP synthesized and secreted by Sertoli cells in the testis is transported to the epididymal duct via testicular fluid and is taken up by epithelial cells of the proximal segments.
mesh_terms: D000596:Amino Acids / Q000032:analysis; D000727:Androgen-Binding Protein / Q000302:isolation & purification*; D000818:Animals; D002241:Carbohydrates / Q000032:analysis; D002352:Carrier Proteins / Q000302:isolation & purification*; D004822:Epididymis / Q000032:analysis*; D007106:Immune Sera; D007118:Immunoassay; D046911:Macromolecular Substances; D008297:Male; D008970:Molecular Weight; D011817:Rabbits / Q000276:immunology; D051381:Rats; D013737:Testis / Q000032:analysis
chemical_list: D000596:Amino Acids; D000727:Androgen-Binding Protein; D002241:Carbohydrates; D002352:Carrier Proteins; D007106:Immune Sera; D046911:Macromolecular Substances