Abstract for Grants
The X-ray core at Beckman Research Institute of City of Hope is a state-of-the-art crystallization and X-ray facility for basic and translational science. To facilitate investigators, the core also provides protein expression and purification as well as numerous biophysical methods to characterize stability, oligomerizaiton, and kinetic and thermodynamic properties.This facility houses a Mosquito Crystallization robot that uses 50 to 100 nL volumes and permits three different crystallization formats; hanging drop, sitting drops or batch methods under oil. The facility has multiple commercial crystallization factorials (e.g., Hampton, Jena, Nextal), To follow crystallization trials, two Formulatrix Imaging Robots at different temperatures are used that automatically image each drop, according to a preset schedule (e.g., nightly). This allows follow-up of crystallization trends at two temperatures and produces a visual record for analysis. A Rigaku 007HF generator, R-axis IV++ imager and an Oxford cryojet (for data collection at 100 K) is used for in-house diffraction studies and access to SSRL and other synchrotrons is available The facility has all the software and computational hardware necessary to determine structures of macromolecular complexes.
In addition, our core facility has capabilities to express and purify large quantities of high quality protein (1 to 100 mg +) using a variety of expression systems (bacteria, insect and mammalian cells). Biophysical characterization of macromolecules and small molecules can be carried out using SPR, ITC, AUC, analytical SEC, KinExa and CD and fluorescent spectroscopy.