Reduced Dimensionality NMR Spectroscopy for Structural Genomics

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SzyperskiMCB 0075773This research focusses on the development and application of novel Bio-NMR techniques which promise to have a broad imact for science infrastructure beyond use in structural genomics.NMR is expected to play an important role to harvest the fruits of genome sequencing projects by supporting the functional analysis of gene products in "structural genomics". The mass production of protein structures in the framework of this "big science" project requires new methodology for cost-effective structure determination. This research is dedicated to further develop reduced-dimensionality (RD) triple resonance NMR experiments for use in "structural genomics". Such spectroscopy promises to be ideally suited because it allows, firstly, matching of the NMR measurement time to sensitivity requirements for signal detection while retaining a high digital resolution for automated spectral analysis, and secondly, assigning proteins with a smaller set of NMR spectra when compared with conventional spectroscopy. The current proposal invokes both fundamental research, i.e., Bio-NMR methods development, and applied research targeting the commercial effeciency of NMR parks or consortia operated for structural genomics projects. Research will aim at the development of a protocol that allows identification of minimal sets of NMR spectra for structure determination of a given protein in a structural genomics enterprise. Considering RD NMR experiments, this will allow one retaining a high digital resolution when investments of spectrometer time have to be minimized, as it can be expected within the frameworkk of a structural genomics project. Moreover, for application in structural genomics, the impact of RD spectroscopy for the automatic resonance assignment of proteins will be evaluated, and an 'RD NMR package' comprising RD pulse programs, NMR parameter sets and job-files for automated spectral analysis will be devised. Moreover, research will focus on the implementation of improved RD experiments including a suite of schemes that allows measurement of residual dipolar couplings in dilute liquid crystalline media. This development opens an avenue to merge their measurement with the resonance assignment, as well as to make use of variations of sums of scalar and residual dipolar couplings along the polypeptide chain to establish sequential connectivities.This research offers outstanding opportunities for involving students at all stages in their academic development, and can be expected to play a valuable role in integrating research and education.