Biological NMR for Challenging Biomedical Problems. (360G-Wellcome-104920_Z_14_Z)

Funds are requested to purchase a new generation of NMR console and cryogenically cooled probe for our outdated 750MHz NMR Spectrometer (funded in 2001 by WT) which has reached the end of its useful lifetime. Our current instrument is inefficient, unreliable and incapable of modern NMR experiments. These purchases will transform NMR capability in the Astbury Centre for challenging biomedical projects that require the sensitivity, artefact suppression and stability only possible with modern elect ronics. For 443k of WT funds, and University investment of 647k, the new equipment will enable cutting edge biological NMR techniques, including: 1) analysis of protein dynamics (via relaxation experiments); 2) structural characterization of 'invisible', lowly-populated protein conformations (via relaxation dispersion experiments); 3) characterization of large supramolecular assemblies and membrane proteins (via multidimensional methyl NMR); 4) time-efficient structural calculations of small (20kDa) proteins (structure in 1 week); 5) real-time kinetic measurements of folding and enzymatic activity (via rapid acquisition techniques); 6) analysis of unstable samples (via fast techniques allowing data acquisition up to 100 time faster) and materials available only at low concentration (10-100 uM); 7) analysis of samples in biological buffers (up to 1M salt) 8) time-efficient ligand screening (via rapid data acquisition and sample changer).