STRUCTURAL STUDIES OF THE DNA-BINDING AND LIGAND-BINDING DOMAINS OF THE HUMAN

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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. Since Spring 2005 we collected structural data at the APS which related to ERalpha ligand-binding domain (LBD) complexed with novel ligands that vary in their potency and selectivity for ERalpha and ERbeta. From these data, we were able to speculate on a possible mode of binding of these compounds to ERb for which they are more selective in solution. These OBCP compounds with their novel 3D scaffolds are being evaluated for various applications since their biological effects from being full agonists, partial agonists/antagonists, or complete antagonists, depending on the tissue and target gene promoter context. We continue our structure-function characterization of the two known ER subtypes with crystal complexes of ERbeta LBD and the OBCP compounds. These structures will further our understanding of how to better use the OBCP scaffold for the development of improved subtype selective agonists and/or novel antagonists. Another important development is that we finally have diffraction quality crystals of an ERbeta truncation mutant that includes the DNA-binding domain (DBD), a hinge region and the following LBD. This promises to be a high resolution structure since it diffracted to 1.34A when we tested it just before the end of the previous run. This is an exciting construct because, to date, there are no available structures of ERalpha/beta that include both the DBD and LBD. Determining this structure will bring us closer to understanding the structure-function regulation of the entire ERbeta protein, a therapeutically important molecule with multiple domains.