600 MHz NMR Spectrometer for Structure Determination of Proteins and RNA

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The Biochemistry Division in the Department of Chemistry and Biochemistry at the University of Colorado, Boulder will purchase a state-of-the-art four channel 600 MHz NMR spectrometer with partial support from the NSF Multi-User Biological Equipment program. This instrument is required to perform multi-nuclear multi-dimensional NMR experiments used in the solution structure determinations of isotopically labeled proteins and RNAs. Triple-resonance multi-dimensional NMR techniques now allow complete through bond resonance assignment of proteins which means that the primary bottleneck in solution structure determinations of the proteins (the assignment of the resonances) can now be made much more unambiguously and in much less time than previous techniques. The modern NM R solution structure determinations of a protein requires isotopically labeled protein and access to a triple-resonance instrument that has pulse shaping and pulse-field gradient capabilities. The multi-nuclear and multi-dimensional NMR techniques that have proved so successful in solution structure determinations of proteins can now be applied to structural studies of RNAs and protein-RNA interactions. This additional NMR instrumentation is required to meet the demand for NMR time arising from the recent expansion of the faculty (Dr. Wuttke) in the Biochemistry Divisimn. The requested instrumentation will vastly improve the NMR capabilities and will accelerate research in the following areas: - Solution Structural Studies of RNA and Protein-RNA Systems (Pardi) - Development of NMR Techniques for Solution Structure Determinations of RNA (Pardi) - NMR Structural Studies of Cardiac Troponin C (Falke) - Structural Studies of the Active Site of the Group I Ribozyme (Cech) - Structure and Function of Catalytic RNAs (Uhlenbeck) - Solution Structural Studies of the trans Acting Viral Proteins Tax, pX and Their Complexes (Wuttke)