Contact Information
Core Director: Larry J. DeLucas, OD, PhD, DSc (Hon.)
Email: Delucas@cbse.uab.edu
Phone: 205-934-5329
Web site: http://www.cbse.uab.edu/
Center Association: Center for Biophysical Sciences and Engineering (CBSE), Comprehensive Cancer Center (CCC)
Established: 1975
Mission
The X-Ray Crystallography Shared Facility provides CCC members, UAB investigators, and regional scientists access to a premiere facility for protein crystal structure determinations for both aqueous and membrane proteins. The facility includes automated high throughput systems for crystallization screening capable of preparing up to 10,000 crystallization experiments a day in nanoliter to microliter volumes as well as other novel techniques to optimize crystal growth in order to obtain the diffraction quality crystals necessary for structure determination. The CBSE facility maintains four operational bays for in-house diffraction data collection along with access to two dedicated synchrotron beamlines at the Argonne National Laboratory. Our facility positions researchers with competitive advantages to rapidly determine protein structures of interest.
Facility Description
The facility is located in The Center for Biophysical Sciences and Engineering building and operated under the direction of Dr. Larry DeLucas, with five core personnel and three support staff. Its four service areas are as follows:
1) High Throughput (HTP) protein crystallization and Co-Crystallization screening facility. This 680 square foot facility utilizes the following integrated platform of systems:
· A HTP automated system for rapid generation and optimization of specialized solutions used in crystallization screens (Recipe Maker™).
· A new (2008) Rigaku Phoenix RE Liquid Handling System for high throughput low volume protein crystallization experimentation.
· Two automated HTP liquid dispensing systems for submicroliter volumes (the custom built NanoScreen™ and the commercial Cartesian HoneyBee™ System).
· An automated microchip system for free interface diffusion crystallization along with an automated imaging station for microchips (a.k.a. the Fluidigm Topaz Microchip System™).
· An automated HTP capillary counter diffusion system (the HoneyComb by Genomic Solutions™).
· A HTP automated system for optical recognition and scoring of crystals (Crystal Score™).
· A dynamically controlled crystal growth kinetic system for optimization of crystal growth (Vapor Pro™).
· Use of a virtual screening program (utilizing predictive algorithms) for the optimization of crystallization conditions, thus reducing amount of sample and number of experiments needed to define high quality crystal growth. This technology supports efforts to find new or optimize present conditions that improve crystal growth for both soluble and membrane proteins.
· HTP Self Interaction Chromatography, a customized HPLC system for accurate protein-protein interaction experiments for the determination of second virial coefficient measurements of proteins. Improved crystallization parameters for proteins as well as optimizing solutions for the stabilization of protein pharmaceutical formulations represents examples of protein solution phenomena that can be measured with thermodynamics measurements of protein-protein interactions. We can measure PPI through the determination of the second virial coefficient. The CBSE has developed a - HTP analytical method and automated system that allows users to quickly determine optimal solutions.
2) Synchrotron X-ray source. Since 1997, UAB has been a member of the Southeast Regional Collaborative Access Team (SERCAT) with dedicated access to two of the most powerful beamlines at the Argonne National Laboratory in Chicago. This access provides 15 or more days of dedicated access for UAB at the most powerful synchrotron facility in the U.S. By using the multiple wavelengths (only available at synchrotrons), the structure of a large protein can be determined in one week or less with suitable crystals. Frozen crystal samples can also be shipped to SER-CAT and diffraction data can be collected utilizing the SERCAT “Mail-in Crystallography†program.
3) Laboratory X-ray source. This 2,116 square foot facility maintains four fully operational bays for in-house X-ray diffraction experiments. The CBSE has 12 PhD level crystallographers in house.
4) Crystallography Computing. This 646 square foot computing facility provides a central computing network system for data processing, data transfer, structure determination, graphics, and database management in conjunction with all the latest software modeling programs available. Remote access to a UAB 128-Beowulf Intel® processor-based Linux cluster allows “pipeline†approaches to be used to automate and accelerate the structure determination process.
Users of this facility can also contact the CBSE for information on our Biomolecular Physics laboratory and the availability of these complimentary techniques used by researchers to support crystallization and compound development. The CBSE Biomolecular Analysis Group currently uses biocalorimetry data and other biophysical tools to enhance the design and optimization of lead compounds. Isothermal Titration Calorimetry and Differential Scanning Calorimetry can advance your research with applications in: 1) protein engineering, detecting misfolded domains; 2) proteomics, energetic domain architecture; 3) optimization of crystallization conditions, 4) biopharmaceutical formulations; 5) drug discovery: developing universal assay for lead identification, development; and 6) understanding the importance of functional groups, salvation, hydrophobicity and conformation.
Research Information
The facility provides structural biology support from cloning and expression through structure determination and combinatorial chemistry for researchers interested in macromolecular structure-function relationships and drug discovery via intelligent or structure-based combinatorial design. Protein structural information is used for the discovery and synthesis of complimentary compounds that augment or inhibit the protein's biological activity for drug discovery applications. Current drug development programs supported by the facility focus on cancer, infectious diseases, cardiovascular disease, bacterial infections, and immune response. The CBSE has a particular interest in crystallizing and solving membrane protein structures.
Approved by: Larry J. DeLucas, OD, PhD, DSc (Hon.), Director
Date: March 18, 2008
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