Discovery of novel protein complexes across human gut bacteria

Division Of Basic Sciences - Nci

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Abstract

We have begun to generate whole-cell protein-protein interaction maps of the pathogen Salmonella enterica. We are using proteome fractionation that we are subjecting to co-fractionation MS, crosslinking MS and evolutionary contact analysis. This will combine into a deep interactome with residue-level structural resolution. This project is still in the early stages so does not have many results to report. In parallel, we are using metabolically incorporated photo-amino acids containing diazirine groups crosslink to nearby residues when they are activated with UV-light. We are using Metabolic incorporation to stochastically incorporate photoLeucine at leucine sites. The leucine T-RNA synthase will accept this. We have managed to get the incorporation rate up to over 5%. Here we have some preliminary data from the salmonella proteome where we have mapped the photoleucine crosslinks onto some AlphaFold2 models.