Innovative protein crystallization screens

Crystallization Screens Crystallization Optimization Cryo Screens Phasing

Phase knowledge enables rational screens for protein crystallization.

We show that knowledge of the phase behavior of a protein allows one to create a rational screen that increases the success rate of crystallizing challenging proteins. The strategy is based on using microfluidics to perform large numbers of protein solubility experiments across many different chemical conditions to identify reagents for crystallization experiments. Phase diagrams were generated...

Crystallization data mining in structural genomics: using positive and negative results to optimize protein crystallization screens.

Recent efforts to collect and mine crystallization data from structural genomics (SG) consortia have led to the identification of minimal screens and novel screening strategies that can be used to streamline the crystallization process. Two groups, the Joint Center for Structural Genomics and the University of Toronto, carried out large-scale crystallization trials on different sets of bacteria...

Coupling of MBP fusion protein cleavage with sparse matrix crystallization screens to overcome problematic protein solubility.

Nuclear magnetic resonance (NMR) and crystallographic techniques require large quantities of pure, soluble protein. However, numerous target proteins exhibit limited solubility resulting in low recovery and reduced concentrations following purification. Molecular biological techniques have been developed to assist in this area including the use of a fusion protein attached to the N or C terminu...

Protein crystallization screens developed at the MRC Laboratory of Molecular Biology

In order to solve increasingly challenging protein structures with crystallography, crystallization reagents and screen formulations are regularly investigated. Here, we briefly describe 96-condition screens developed at the MRC Laboratory of Molecular Biology: the LMB sparse matrix screen, Pi incomplete factorial screens, the MORPHEUS grid screens and the ANGSTROM optimization screen. In this ...