The X-Ray Crystallography Center in the Department of Structural Biology is a shared resource designed to facilitate studies of protein and macromolecular structure and function by investigators at St. Jude Children’s Research Hospital. The center is equipped with numerous instruments supporting every aspect of macromolecular crystallography.
Three crystallization robots offer rapid screening at different temperatures using commercially available kits and nanoliter protein volumes. Most notably, the versatile Mosquito LCP from TTP Labtech allows for crystallization of membrane proteins using lipidic mesophases as well as protein crystallization by traditional vapor diffusion methods. The Dragonfly (TTP Labtech) liquid handler aids in custom screen formulation for crystal optimization. Dedicated incubation rooms for 18 °C and 4 °C crystal plate storage and visualization are available. In addition, we have a 1,000-plate capacity storage hotel from Formulatrix that greatly simplifies the task of visual assessment of crystallization results and protein vs. salt discrimination; the Rock Imager 1000 incubates crystal plates and is automated for high-quality visible and UV image capture.
Our X-ray laboratory is well equipped with a complete Bruker AXS D8 Venture X-ray diffraction system, and St. Jude is a member of the South East Regional Collaborative Access Team (SER-CAT) at the Advanced Photon Source (APS). The extremely intense and tunable synchrotron-radiation available at the SER-CAT 22-ID and 22-BM beamlines have allowed St. Jude researchers to obtain numerous high-quality and novel structures. The insertion device (ID) beamline features microbeam focusing for improved diffraction from small crystals and a high-speed detector for fast data acquisition (~3-10 minutes per crystal). We have convenient and regular use of the synchrotron, remote data collection is routine, and the hospital’s three shares in SER-CAT yield about 20 days of data collection time per year.
In addition, our extensive computing infrastructure supports all aspects of the structure determination process and post-structure analysis.