title: Heterotetrameric structure of the human progesterone receptor.
abstract: Nonactivated progesterone receptors in extracts of human T47D mammary carcinoma cells were investigated. Chemical cross-linking with dimethyl suberimidate resulted in complete stabilization of the A and B receptors with an average molecular mass of 340 kDa. For analyzing the subunit structure, we concentrated on the larger B receptor, which was separated from the A form by immunoaffinity chromatography. Progressive cross-linking of the photoaffinity-labeled receptor resulted in patterns of labeled bands in SDS gels, which are indicative of a heterotetrameric structure. It consists of one receptor polypeptide in association with two 90-kDa subunits and one polypeptide of approximately 60 kDa. The completely cross-linked B receptor has a molecular mass of approximately 390 kDa. To identify the subunits, the oligomeric B receptor was cross-linked with a cleavable bisimidate, highly purified by immunoaffinity chromatography, and analyzed by gel electrophoresis and immunoblotting. The receptor polypeptide has a mass of 116.5 kDa. The 90-kDa band was identified as the heat shock protein hsp90 and was roughly twice as intense as the receptor polypeptide. By use of specific antibodies, we identified the fourth receptor subunit as a 59-kDa protein (p59); we did not obtain any evidence for the heat shock protein hsp70 being a receptor component. We suggest an analogous heterotetrameric structure for the nonactivated A receptor.
authors.affiliation: Institut für Biologische Chemie, Universität Heidelberg, Federal Republic of Germany.
authors.forename: P
authors.initials: P
authors.lastname: Rehberger
authors.forename: M
authors.initials: M
authors.lastname: Rexin
authors.forename: U
authors.initials: U
authors.lastname: Gehring
mesh_terms: D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D003432:Cross-Linking Reagents; D006801:Humans; D066298:In Vitro Techniques; D046911:Macromolecular Substances; D008970:Molecular Weight; D011980:Receptors, Progesterone / Q000737:chemistry / Q000648:ultrastructure*; D014407:Tumor Cells, Cultured
chemical_list: D003432:Cross-Linking Reagents; D046911:Macromolecular Substances; D011980:Receptors, Progesterone