The Structural Genomics of Pathogenic Protozoa (SGPP) consortium aims to achieve high-throughput determination of protein structures from pathogenic protozoa of major global medical relevance. These include several species of Trypanosomatids, responsible for sleeping sickness, Chagas' disease, and cutaneous, visceral and mucocutaneous leishmaniasis; and Plasmodium falciparum, the causative agent of lethal malaria with several hundred million cases annually and one to two million deaths, mainly children. These organisms are only distantly related to other species and characterized by many unique biological features.
We aim to determine a total of NN soluble, heteromultimeric and membrane proteins. Special characteristics of the proposed research are: 1. Using protein structure prediction methods and medical relevance in target selection; 2. Rapid evaluation of expression levels and solubility of thousands of variants of proteins obtained by scrambled sequences from different strains; 3. Using two-hybrid methods to discover and solve structures of soluble heteromultimers; 4. Significant emphasis on membrane proteins with discovery, by two-hybrid methods and Fv phage libraria,of interacting soluble partner proteins to be used for co-crystallization and MAD phasing; 5. Intensive use of robotics in the crystallization, crystal mounting, data collection steps; 6. Development of bromine-containing co-crystallant libraries; 7. Exploring the power of crystal annealing in improving mosaicity and resolution; 8. Using predicted protein structures in electron density interpretation; 9. Using ligand docking procedures, deep sequence family alignments and very weak structural homologies to derive function from structure; 10. Create a SGPP website and a SGPP relational data base by which protocols for protein expression, purification, crystallization and structure determination, as well as coordinates and structure factors are made available to the public.
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