title: Transformation of the estrogen receptor detected by two monoclonal antibodies.
abstract: The estrogen receptor from fetal guinea-pig uterus is recognised by two monoclonal antibodies (H222 and H226) developed against the human estrogen receptor but it interacts differently with each of them. The H222 antibody, whose epitope is located in the hormone-binding domain of the receptor, shifts the sedimentation coefficient of the nonactivated oligomeric receptor in low salt sucrose gradients from 9S to 11S. When this oligomeric receptor-H222 complex is centrifuged in high salt gradients, it dissociates to an 8S monomer-H222 complex, indicating that all the estradiol-binding units present in the nonactivated receptor can bind the H222 antibody. In contrast, the H226 antibody, whose epitope is located close to the DNA-binding domain, shifts the sedimentation coefficient of the nonactivated receptor only to 9.4S and when this complex sediments in high salt gradients, it dissociates to a 7S monomer-H226 complex plus a 4.5S monomeric receptor not bound to the antibody. This observation suggests that not all the H226 epitopes are accessible in the nonactivated receptor. On the other hand, the temperature-activated receptor reacts with the H226 antibody to form two complexes sedimenting at 7S and 9S in high salt gradients. This 9S complex indicates the formation of a homodimer that binds two molecules of the H226 antibody. However, only one H222 epitope seems to be accessible in this dimeric form of the receptor, since only one 8S complex is observed when the activated receptor reacts with the H222 antibody. In addition, binding to the H222 antibody before activation prevents the dimerisation. This suggests that the H222 epitope is near or directly involved in the dimerisation domain. Interaction of the H222 and H226 antibodies with the estrogen receptor reveals modifications of its structure during activation, and consequently of the exposure of its functional domains.
authors: C.N.R.S. Steroid Hormone Research Unit, Foundation for Hormone Research, Paris, France.
authors: N
authors: N
authors: Giambiagi
authors: J R
authors: JR
authors: Pasqualini
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal; D002467:Cell Nucleus / Q000378:metabolism; D000939:Epitopes / Q000032:analysis*; D005260:Female; D005333:Fetus; D006168:Guinea Pigs; D006358:Hot Temperature; D007700:Kinetics; D046911:Macromolecular Substances; D011247:Pregnancy; D011960:Receptors, Estrogen / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D014599:Uterus / Q000378:metabolism*
chemical_list: D000911:Antibodies, Monoclonal; D000939:Epitopes; D046911:Macromolecular Substances; D011960:Receptors, Estrogen