X-ray Crystallography
Scientific Objectives: The unique information derived from protein structure determination acts as an accelerant for subsequent biochemical studies. Thus, collaborating scientists create a cycle of structure determination and biochemistry that pushes their research forward. There exists a similar relationship with the field of computer-aided drug design (CADD), because the structural techniques are the most direct way to assess potential new drugs at the molecular level and subsequently improve there efficacy. The x-ray crystallography shared service will be most valuable in helping the Cancer Center research community bring into focus the molecular bases of defects in the machines that govern our cellular well-being and the development of drugs that combat a variety of diseases.
Personnel:
Xiapeng Hu, Ph.D. Dr. Hu currently coordinates the activities of the X-ray Crystallography Shared Resource. Dr. Hu has seven years of experience as an x-ray crystallographer and is familiar with maintenance of x-ray equipment. As the manager of the x-ray shared service facility, Dr. Hu is responsible for both the administrative and technical operation of the resources provided by the facility. More specifically, he determines the schedule for use of the x-ray equipment, bills users for services rendered, and ensures that the equipment, including computers and their associated software packages, is properly maintained. He is also responsible for training new users on the x-ray equipment and assisting them in the structure solution process from the beginning (i.e obtaining crystals) to the end (i.e. a final model worthy of publication). He will be responsible for any contract work performed by the facility. Dr. Hu will also attend all trips to synchrotron facilities for experiments beyond the capabilities of our in-house equipment, thereby acting as a representative of the facility. This will ensure that operators at synchrotron x-ray stations will become familiar with our work and provides a knowledgeable person on each trip to prevent lost productivity that often occurs when novice users are faced with an unfamiliar workstation. Finally, he will ensure that radiation safety guidelines are strictly enforced at all times. Dr. Hu will thus serve as the primary interface between the x-ray facility and the scientific community at the Greenebaum Cancer Center, helping to guide users through the often daunting steps that turn crystals into a finished structure that helps to answer pressing scientific questions.
The decisions regarding the day-to-day operations of the x-ray crystallography shared resource are made by an internal oversight committee comprised of Drs. Eric Toth, Averell Gnatt, and Adrian Batchelor. This committee meets monthly with the facility manager Dr. Hu present to discuss pressing issues pertaining to the shared resource. Their qualifications and distinct roles (in addition to their collaborative efforts on devising an operational strategy for the shared resource) are detailed below:
Eric A. Toth, Ph.D. Dr. Toth is an Assistant Professor in the Department of Biochemistry and Molecular Biology and a member of the Cancer Center’s Molecular and Structural Biology program. He has worked in the field of protein crystallography for 12 years on a variety of protein systems, including adenylosuccinate lyases, DNA polymerases, and helicases. Dr. Toth has extensive experience in de novo protein structure determination from both data collected using standard in-house diffraction equipment and cutting edge technologies used at synchrotron facilities. Dr. Toth also has a keen interest in the computational aspects of protein crystallography and has therefore been charged with designing and overseeing the computational infrastructure of the shared resource. To ensure that adequate disk space for all users is available and easy to manage, the shared resource purchased a 4 terabyte file server, which is currently managed by Dr. Mark Pohl, the director of the University’s Bioinformatics Core Facility.
Averell L. Gnatt, Ph.D, Dr. Gnatt is an Assistant Professor in the Departments of Pharmacology and Experimental Therapeutics and Pathology and a member of the Cancer Center’s Molecular and Structural Biology program. In addition to his training in neurobiology, Dr. Gnatt has 13 years of experience as a protein crystallographer. His focus has been primarily on the structure of the eukaryotic transcription machinery. Because of the enormity of the RNA Polymerase II complexes that Dr. Gnatt studies, he has extensive experience with data collection problems posed by large macromolecular complexes with regards to in-house equipment and synchrotron facilities.
Adrian H. Batchelor, Ph.D. Dr. Batchelor is an Assistant Professor in the Department of Pharmaceutical Sciences in the University of Maryland School of Pharmacy. He has 11 years of experience in protein crystallography, working on transcription factor-DNA complexes and currently structure determination and functional analysis of malaria parasite surface proteins. Dr. Batchelor oversaw the installation of our x-ray diffraction equipment and undertook the operation and maintenance of the equipment from the time of its installation. He is well-versed in the inner workings of the x-ray source, detector, and cryocooling apparatus and often advises Dr. Hu on routine maintenance. Because his lab is adjacent to the x-ray facility, Dr. Batchelor also performs upkeep of the system when Dr. Hu is unavailable to do so.
Services:
The X-ray Crystallography Shared Service provides the following services to investigators:
* protein crystallization
* X-ray characterization of crystals
* data collection
* processing and quality analysis of data
* structure determination
* molecular modeling
* molecular docking and structure visualization and analysis.
Instrumentation:
* Rigaku-MSC Micromax 7 generator equipped with a Raxis-4++ image plate detector
* State-of-the-art detector systems
* Oxford Cryosystems Cryocooler
* Crystal manipulation accessories, such as microscope with attached camera for crystal viewing/documentation
* Computer suite for data analysis and structure determination
Popular x-ray diffraction data processing software suites, such as HKL2000, CCP4, CNS, and O.
Pricing:
Use of the Facility for authorized personnel $50/hour; $450 maximum per day
Training by our staff $50/hour (one hour minimum)
Crystallization Screening $250 (typically for 48 conditions)
Optimization of Crystal Growth $50/hour + materials
Crystal Diffraction Testing $50/hour
X-ray Data Collection Same as use of the facility
Data analysis $50/hour
Protein Purification Contact us
Other Services Contact us
Contact Information:
xhu@umaryland.edu HSF II, room 516
Copyright © University of Maryland School of Medicine
Comments: 0