Emerald BioStructures, Inc. Is developing an integrated set of crystallization research tools and software that are designed for high-throughput crystallization by robots as well as the standard laboratory use by individual researchers with little or no prior crystallization research experience. These tools provide a streamlined and systematic approach for identifying crystallization conditions that yield X-ray diffraction quality protein crystals. In this application, the investigators propose to develop a systematic and generally applicable method for the preparation of 10s of milligrams of purified proteins that will fed directly into their integrated crystallization package, thereby specifically addressing the rate-limiting aspects of the Structural Genomics Initiative. We propose to fuse the complementary expertise of Molecular Simulations, Inc. (Protein structure analysis and target selection using GeneAtlas software), Invitrogen Corporation (construction and validation of expression-ready Gene Storm vectors) and, with those of Emerald BioStructure, Inc. (Crystallization reagents and software, and X-ray structure determination). Our goal is to automate the production and crystallization of full-length target proteins of unknown structure or function. This is made possible by the availability of their novel Crystal Monitor software which enables high speed collection of crystallization trial data using speech recognition, which will be programmed to drive custom built crystallization robots.We will take advantage of Invitrogen's ambitious efforts to generate expression ready GeneStorm vectors encoding all human open reading frames (ORF's). Molecular Simulations, Inc., will employ their GeneAtlas software to identify target human ORFs whose structure determinations are expected to yield completely novel structural information. Together the companies will explore the utility of systematic procedures for producing, purifying, and crystallizing soluble native or refolded proteins expressed in E. Coli or insect cells. In addition, they propose an innovative procedure for renaturing and crystallizing proteins in the same vapor diffusion chamber. Crystallization of one or more novel full-length proteins will establish the feasibility of their approach, thereby enabling a more fully automated Structural Genomics effort in subsequent phases of research. PROPOSED COMMERCIAL APPLICATION: If successful, this Phase I project would form the basis for an automated system for protein crystallization. Hence, the proposed research will facilitate the high- throughput X-ray structure determination of proteins, and help to spawn a new service industry in macromolecular crystallography.