Protein engineering to transform reliability of protein crystallization

While X-ray crystal structures of proteins are terrifically informative, both in basic research (6 Nobel prizes in 25 years) and drug design, the major stumbling block remains the need to obtain crystals of the biological sample (protein, protein complex, virus) suitable for X-ray analysis.  Even for a biochemically well-behaved sample, the likelihood of it crystallizing is always low, and there are no rules or reliable protocols to improve this.  Structure-based drug discovery similarly requires diverse crystal forms to be available for the protein; this too cannot currently be easily induced.

This project addresses the pressing need for rapid and simple protein engineering techniques to make samples crystallize routinely.  This will entail, depending on the student's background, skills and preferences:

• Develop streamlined protocols for parallel generation of large numbers of crystallization chaperones, e.g. using in vivo selection directly coupled to over-expression, with DNA libraries of binding scaffolds;
• Evolve binders that favour crystallization, by adapting existing motifs through iterative design supported by high-throughput crystallization, e.g. Gluebodies [Ye et al, 2024]; 
• Adapt protein design algorithms to find surface mutations that better allow the protein to pack into crystals, e.g. by machine learning of crystal contacts in the PDB;
• Streamline protocols for rapid large-volume parallel expression, purification and crystallization of large numbers of diverse protein variants; 
• Develop generic scaffolds that crystallize robustly, and ways of tethering target proteins to the scaffold.