CONFORMATIONAL CHANGES AND MODE OF BINDING OF AGONIST TO ESTROGEN RECEPTOR

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This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. David Budil requested us to characterize structural response of the estrogen receptor ligand binding domain (ER-LBD) to a variety of ligands ranging from strong estrogens to strong antiestrogens using electron spin labeling. The technical aims for the initial period involved developing site-directed spin-labeled mutants of the ER-LBD and synthesizing new spin-labeled ligands for the proposed studies. Budil's research group has optimized cell culture production of ER-LBD and expressed a number of single and doubly labeled mutants. In addition a new spin label was synthesized for attachment to site-selected cysteine residues in this protein and substantial progress was made towards the synthesis of spin-labeled ligands with a range of agonist/antagonist activities. Initial electron spin resonance studies have established that the selected label locations will be acceptably sensitive to the type of ligand present.