Collaborative Research: RUI: Expanding the repertoire of chemical tools to study and characterize bacterial Gcn5-related N-acetyltransferase functions

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This National Science Foundation Research in Undergraduate Institutions award from the Chemistry of Life Processes Program funds Dr. Misty L. Kuhn from San Francisco State University (SFSU) and Dr. Daniel P. Becker from Loyola University Chicago to create biochemical tools to study enzymes in the family of Gcn5-related N-acetyltransferase (GNAT) proteins of unknown function from the soil microorganism, Pseudomonas aeruginosa. Specifically, these tools are used in combination with structure determinations via X-ray crystallography to gain a greater understanding of how molecules bind to GNATs. Since relatively little is known about the structure/function relationship of GNATs from P. aeruginosa, this work fills this gap in knowledge. Determining these functions is important because human welfare depends on healthy agriculture. This project enhances the research infrastructure at SFSU by establishing its first protein crystallization resource and increasing the repertoire of sophisticated biochemistry skills of underrepresented minority and disadvantaged undergraduate students. The goal of this project is to improve the functional annotation of P. aeruginosa GNATs of unknown function using a combination of protein X-ray crystallography, molecular modeling, organic synthesis, and enzymology. The architecture of the acceptor site of several GNAT proteins are probed using biochemical tools and X-ray crystallography. Active site residues that are critical for catalysis and substrate specificity are identified and tested using molecular modeling, site-directed mutagenesis, and enzyme kinetics. The results of this project provides the foundational knowledge to categorize experimentally uncharacterized members of the GNAT family and improve computational functional annotation of GNATs of unknown function.