Journal article
Molecular insights into the structure and function of the Staphylococcus aureus fatty acid kinase
The Journal of biological chemistry, Vol.300(12), 107920
12/2024
DOI: 10.1016/j.jbc.2024.107920
PMCID: PMC11617999
PMID: 39454961
Abstract
Gram-positive bacteria utilize a Fatty Acid Kinase (FAK) complex to harvest fatty acids from the environment. This complex consists of the fatty acid kinase, FakA, and an acyl carrier protein, FakB, and is known to impact virulence and disease outcomes. Despite some recent studies, there remains many outstanding questions as to the enzymatic mechanism and structure of FAK . To better address this gap in knowledge, we used a combination of modeling, biochemical, and cell-based approaches to build on prior proposed models and identify critical details of FAK activity. Using bio-layer interferometry, we demonstrated nanomolar affinity between FakA and FakB that also indicates that FakA is dimer when binding FakB. Additionally, targeted mutagenesis of the FakA Middle domain demonstrates it possesses a metal binding pocket that is critical for FakA dimer stability and FAK function in vitro and in vivo. Lastly, we solved structures of the apo and ligand-bound FakA kinase domain to capture the molecular changes in the protein following ATP binding and hydrolysis. Together, these data provide critical insight into the structure and function of the FAK complex which is essential for understanding its mechanism.
Details
- Title: Subtitle
- Molecular insights into the structure and function of the Staphylococcus aureus fatty acid kinase
- Creators
- Megan J. Myers - University of Kansas Medical CenterZhen Xu - University of IowaBenjamin J. Ryan - University of Kansas Medical CenterZachary R. DeMars - University of Kansas Medical CenterMiranda J. Ridder - University of Kansas Medical CenterDavid K. Johnson - University of KansasChristina N. Krute - University of Kansas Medical CenterTony S. Flynn - University of Kansas Medical CenterMaithri M. Kashipathy - University of KansasKevin P. Battaile - New York Structural Biology CenterNicholas Schnicker - University of IowaScott Lovell - University of KansasBret D. Freudenthal - University of Kansas Medical CenterJeffrey L. Bose - University of Kansas Medical Center
- Resource Type
- Journal article
- Publication Details
- The Journal of biological chemistry, Vol.300(12), 107920
- DOI
- 10.1016/j.jbc.2024.107920
- PMID
- 39454961
- PMCID
- PMC11617999
- NLM abbreviation
- J Biol Chem
- ISSN
- 0021-9258
- eISSN
- 1083-351X
- Publisher
- Elsevier Inc
- Grant note
- Lila Self Graduate Fellowship: AI153773
Funding and additional information-National Institutes of Health grant AI121073 (J. L. B.) , National Institutes of Health grant AI153773 (J. L. B.) , Madison and Lila Self Graduate Fellowship (M. J. M.) , COBRE-PSF NIH P30 GM110761 (University of Kansas) . The content is solely the responsibility of the authors and does not necessarily reflect the official views of the National Institute of General Medical Sciences or the National Institutes of Health.
- Language
- English
- Electronic publication date
- 10/23/2024
- Date published
- 12/2024
- Academic Unit
- Molecular Physiology and Biophysics; Medicine Administration
- Record Identifier
- 9984740950302771
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