Meeting review: 2003 NIH Protein Structure Initiative Workshop in Protein Production and Crystallization for Structural and Functional Genomics.

Abstract

The United States National Institutes of Health (NIH) Protein Structure Initiative (PSI) is a joint government, university, and industry effort, organized and supported by the National Institute of General Medical Sciences (NIGMS), and aimed at reducing the costs in increasing the speed of protein structure determination. Its long-range goal is to make the three-dimensional atomic-level structures of most proteins in nature easily obtainable from knowledge of their corresponding DNA sequences (http://www.nigms.gov/psi). It is the primary U.S. component of a broad international effort in structural genomics, involving at least 20 projects throughout the world. The PSI is now in its fourth year. Nine PSI pilot research centers have been funded to explore the feasibility and impact of genomic scale protein structure analysis. To date, over 500 3D protein structures, providing the first structural representatives for hundreds of protein domain families, have been completed and deposited by the NIH centers into the public Protein Data Bank. In addition, new technologies for protein sample production, data organization, and structure analysis by X-ray crystallography and nuclear magnetic resonance (NMR) spectroscopy have been developed. These technologies increase the efficiency of protein structure determination both for structural genomics and for the broader structural biology community. Although more » progress has been substantial, PSI pilot research centers have identified a number of important bottlenecks that need to be solved to meet the goals of the program. For example, it is now accepted that a major challenge to high-throughput protein structure determination is the fact that for some 70% of targeted proteins, it is difficult to produce protein samples and crystals suitable for structural analysis. In an effort to facilitate an effective exchange of developments and advancements between pilot centers, the NIGMS organized a workshop on gene cloning, protein expression and purification in March 2002. The scope of a subsequent workshop in April 2003 was expanded to include high-throughput methods for protein crystallization. Protein production for structural genomics comprises aspects of target protein selection, cloning and expression of recombinant proteins, cell-culture fermentation, isotopic enrichment with selenium for X-ray crystallography and/or stable isotopes for NMR studies, purification, analytical characterization, growing X-ray quality crystals and the process of organizing these results and reagents into databases and reagent libraries, as well as issues associated with distributing these reagents to the broader community. « less