Structural genomics: keystone for a Human Proteome Project.

Abstract

systematic methods for crystallization screening, cryo-crystallography, undulator sources of synchrotron radiation, charge-coupled device (CCD) detectors, multiwavelength anomalous diffraction (MAD) phasing, and software for automatic generation of protein structures from MAD data2 . The impact of these technologies is just beginning to be realized, but in combination they have the potential for allowing high-throughput analysis of protein structures by X-ray crystallography. Indeed, it has been proposed by some researchers that these combined technologies could be used to design a facility based on one synchrotron beam line with an average throughout approaching one protein structure determination per day. Protein NMR spectroscopy will also play a key role in structural genomics. Although high-resolution NMR structure determinations are currently limited to proteins or protein domains with molecular weights less than ~30 kD, many important structural-genomic targets are in this size range. Information derived from NMR studies, including chemicalshift perturbation data for deriving structure-activity relationships and internal dynamic data derived from nuclear relaxation studies, are complementary to crystallographic data. Solid-state NMR may also have particular value for structural analysis of integral membrane proteins. Recent advances in NMR technologies have the potential to greatly accelerate the process of solution structure determination. These developments include high-level protein production systems for biosynthetic isotope enrichment, triple-resonance methods for determining resonance assignments, combined measurements of NOE, scalar coupling, and residual dipolar coupling data that provide many local and global conformational constraints, software for automated analysis of 3D structures from NMR data, and sensitivity enhancements pro