Macromolecular Structure Seed Grant

Applications accepted for Track 1 (budget of $50,000) or Track 2 (budget of ~$15,000 to $25,000). Applicants with projects focusing on characterizing protein-small molecule interactions should consider applying jointly to Chemoproteomics (See Chemoproteomics/Macromolecular Structure – Joint Seed Grant Proposal)

Macromolecular structure projects will include the production, purification, and biophysical analysis of a protein of interest with the ultimate goal of determining three-dimensional structure using X-ray crystallography.

In-kind support will include one-on-one training, consumables, and access to instrumentation at each stage of the protein production, purification, crystallization, and characterization, including:

• Hands-on protein production using the E. coli recombinant expression system and purification supported by chromatographic methods via Akta Pure Fast Protein Liquid Chromatography (FPLC).
• Biophysical analysis in solution: protein structure stability and oligomerization, thermal/chemical unfolding, and ligand binding affinity measurements through the NanoTemper Prometheus Panta, the Monolith PicoRED/Labelfree, and the new Refeyn TwoMP/MassFluidix instruments.
• Advanced three-dimensional structure methodologies supported by automated liquid handling and crystal imaging, structure solving and refinement, and hands-on X-ray diffraction data processing.

Competitive projects will identify a protein sequence (or mutants of the parent sequence, paralogues, or orthologues). Preferred targets are proteins that are poorly understood and lack sufficient structural data. Proposals will identify one graduate student or postdoctoral scholar who will devote at least 25% of the time to the project. The student or postdoctoral scholar should expect to spend that time in the Nucleus lab space.

Macromolecular Structure is led by Daniel Fernandez (danilo@stanford.edu).