800 MHZ SPECTROMETER FOR BIOMOLECULAR NMR: EAR RESEARCH, DEAFNESS

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This subproject is one of many research subprojects utilizing the resources provided by a Shared Instrumentation 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 grant, which is not necessarily the institution for the investigator. DESCRIPTION (provided by applicant): Seven groups of NMR users from the Columbia, St. Louis, and Kansas City campuses of the University of Missouri and from Washington University-St. Louis seek the first 800 MHz NMR spectrometer for the state of Missouri. The high field spectrometer will introduce structural biology of greater significance to their NIH projects. The University of Missouri-Columbia (UMC), in the geographical center of the life sciences corridor from St. Louis through Kansas City, will host the 800 MHz system. The advisory committee for deciding policy for the 800 will have broad representation from St. Louis to Kansas City. A fee system, uniformly applied among all users, will recover most of the operating costs of the 800. UMC offers a large matching package and new space tailored to hosting the 800 within a new building of its School of Medicine. The funds requested will be applied to an 800 MHz spectrometer with a shielded 18.8 T magnet. A cryogenic probe for outstanding sensitivity remains an additional high priority. The boosts in sensitivity, resolution, and TROSY line sharpening afforded by 800 MHz are needed by all these groups now using 600 MHz equipment. Four groups will employ the 800 for cancer-related work. The Van Doren group will investigate (i) interactions of metalloproteases with inhibitor and substrate proteins and (ii) interactions of transforming tumor viral proteins with their target proteins of host cells. Ma and Deutscher will determine NMR structures of galectin-3 bound to a tumor-targeting peptide and a full-length galectin-3 dimer. Ellen Li's group will investigate how ligands shift the equilibrium dynamics of retinoid-X-receptor (RXR) ligand binding domain. John-Stephen Taylor's group will obtain more detailed structural insights on DNA photoproducts underlying skin cancer. Henzl will study structural features and interactions of principal proteins (OCP2 and OCP1) in the auditory organ of the inner ear, a complex that binds connexin 26 most often mutated in hereditary deafness. Three groups will study DNA-binding protein assemblies. Dupureur's group will undertake the first NMR investigations of dynamics and conformational adjustments of a restriction endonuclease in response to DNA binding and conditions of "star activity". Laity will study structure and dynamics of the zinc-sensing DNA-binding domain of metal-responsive transcription factor-1 that regulates zinc homeostasis in mammals. He will study all six of its zinc fingers free and bound to DNA. Dreyfus and Laity will characterize the structure and dynamics of cytolethal CdtB toxin from pathogenic bacteria, free and bound to DNA. The CdtB subunit is sufficient to cause DNA damage and to arrest the cell cycle in mammalian cells.