The overall goal of the MSGPP Program Project will be to determine crystal structures of biomedically important proteins of pathogenic protozoa in complex with small-molecule ligands. Project 1 will be indispensable to and tightly integrated into the overall Program.
Aim 1 of Project 1 will be to select 300 protein targets from ten pathogenic protozoa (including four Biodefense Category B Priority Pathogens) that will be the focus of all downstream efforts. Genomic data from ten microorganisms will be analyzed for """"""""high-value"""""""" targets that are predicted to be good for drug development. Target selection will also involve input from the outside scientific community. Scientists in Aim 2 will apply computational domain prediction to the proteins so that the expression laboratory (Project 2) can express smaller protein units or variants in order to achieve high success rates for soluble expression and crystallization (Project 3).
In Aim 3, experimental ligand discovery will be carried out before and after protein structures are determined. Strategies to discovering small-molecule ligands will include a) in-house frontal affinity chromatography with diverse compound libraries, b) small-molecule microarray screens (through collaboration), and c) submission of protein targets to NIH molecular screening centers.
Aim 3 will include chemical synthesis capabilities. Compounds that are known to act on the selected targets (so called """"""""piggy-back"""""""" compounds) will be obtained from outside sources.
In Aim 4, in silico ligand discovery will be carried out after the initial structure determination of target proteins. Databases of commercially available drug-like compounds will be docked into all protein pockets of functional significance. The high-scoring ligands will be purchased, assayed for binding, and subsequently included in co-crystallization or """"""""soaking"""""""" experiments. Project 1 will coordinate with the other components of the Program Project through an integrated computer database. There will be continual feedback within the units of Project 1, with the other components of the Program Project, and with outside collaborators to help achieve the Program goals.
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