Molecular basis for the activation of the Fatty Acid Kinase complex of Staphylococcus aureus

Myers, M. J.; Xu, Z.; Ryan, B. J.; DeMars, Z. R.; Ridder, M. J.; Johnson, D. K.; Krute, C. N.; Flynn, T. S.; Kashipathy, M. M.; Battaile, K. P.; Schnicker, N. J.; Lovell, S.; Freudenthal, B. D.; Bose, J. L.

doi:10.1101/2024.03.19.585040

bioRxiv · 10.1101/2024.03.19.585040

Molecular basis for the activation of the Fatty Acid Kinase complex of Staphylococcus aureus

Myers, M. J.·Xu, Z.·Ryan, B. J.·DeMars, Z. R.·Ridder, M. J.·Johnson, D. K.·Krute, C. N.·Flynn, T. S.·Kashipathy, M. M.·Battaile, K. P.·Schnicker, N. J.·Lovell, S.·Freudenthal, B. D.·Bose, J. L.

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Abstract

Gram-positive bacteria utilize a Fatty Acid Kinase (FAK) complex to harvest fatty acids from the environment. The complex, consisting of the fatty acid kinase, FakA, and an acyl carrier protein, FakB, is known to impact virulence and disease outcomes. However, FAKs structure and enzymatic mechanism remain poorly understood. Here, we used a combination of modeling, biochemical, and cell-based approaches to establish critical details of FAK activity. Solved structures of the apo and ligand-bound FakA kinase domain captured the protein state through ATP hydrolysis. Additionally, targeted mutagenesis of an understudied FakA Middle domain identified critical residues within a metal-binding pocket that contribute to FakA dimer stability and protein function. Regarding the complex, we demonstrated nanomolar affinity between FakA and FakB and generated computational models of the complexs quaternary structure. Together, these data provide critical insight into the structure and function of the FAK complex which is essential for understanding its mechanism.

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Myers, M. J., Xu, Z., Ryan, B. J., DeMars, Z. R., Ridder, M. J., Johnson, D. K., Krute, C. N., Flynn, T. S., Kashipathy, M. M., Battaile, K. P., Schnicker, N. J., Lovell, S., Freudenthal, B. D., Bose, J. L.. 2024-03-19. Molecular basis for the activation of the Fatty Acid Kinase complex of Staphylococcus aureus. https://doi.org/10.1101/2024.03.19.585040

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