title: Purification and characterization of a novel intracellular 17 beta-estradiol binding protein in estrogen-resistant New World primate cells.
abstract: Compared to Old World primates, including man, New World primates display target-tissue resistance to gonadal steroid hormones. In female New World primates this resistant phenotype is characterized by elevated concentrations of plasma estradiol and progesterone. Here we describe the discovery of an intracellular estrogen binding protein (IEBP) that acts to concentrate 17 beta-estradiol (E2) in the cytoplasm of New World primate target cells. IEBP was purified by E2-affinity chromatography from postnuclear extracts of the B95-8 cells established from an E2-resistant New World primate. Compared with unpurified extract, affinity-purified IEBP demonstrated a 300-fold enrichment in specific E2 binding activity; half-maximal displacement of [3H]E2 from affinity-purified IEBP was observed with 0.1 nM E2. Affinity-purified extracts were subjected to SDS-PAGE with isolation of a dominant 27-28 kDa protein. N-terminal sequencing of tryptic peptides of the protein showed sequence homology with human heat shock protein-27 (hsp27). By immunoblot and E2 binding capacity, IEBP was 1] 2-3-fold greater in New World than in Old World primate tissues and cell lines, 2] heat-inducible and 3] up-regulated in vivo in the presence of the functioning female gonad. In conclusion, IEBP is a specific E2-interacting heat shock protein in the hsp-27 family that is relatively overexpressed in estrogen-resistant cells.
authors.affiliation: Division of Endocrinology, Diabetes and Metabolism, Cedars-Sinai Burns and Allen Research Institute, UCLA School of Medicine, Los Angeles, California, USA. chenh@cshs.org
authors.forename: Hong
authors.initials: H
authors.lastname: Chen
authors.forename: Bing
authors.initials: B
authors.lastname: Hu
authors.forename: Gang-Hua
authors.initials: GH
authors.lastname: Huang
authors.forename: Amanda G
authors.initials: AG
authors.lastname: Trainor
authors.forename: David H
authors.initials: DH
authors.lastname: Abbott
authors.forename: John S
authors.initials: JS
authors.lastname: Adams
mesh_terms: D000595:Amino Acid Sequence / Q000235:genetics; D000818:Animals; D001667:Binding, Competitive; D002352:Carrier Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D002460:Cell Line; D003593:Cytoplasm / Q000378:metabolism; D004351:Drug Resistance; D004958:Estradiol / Q000378:metabolism*; D004967:Estrogens / Q000494:pharmacology*; D007425:Intracellular Membranes / Q000378:metabolism; D008969:Molecular Sequence Data; D005810:Multigene Family; D009363:Neoplasm Proteins / Q000235:genetics; D011323:Primates; D017386:Sequence Homology, Amino Acid
chemical_list: D002352:Carrier Proteins; D004967:Estrogens; D009363:Neoplasm Proteins; D004958:Estradiol