Domain architecture and catalysis of the Staphylococcus aureus fatty acid kinase

Subramanian, Chitra; Cuypers, M.G.; Radka, C.D.; White, Stephen W.; Rock, Charles O.

doi:10.1016/j.jbc.2022.101993

Cited by 7 publications

(14 citation statements)

References 49 publications

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“…S4). This matches the published C-term FakA structure (PDB 6W6B, 30 ). The FakA C-term structure consists of both β- sheets and α-helices (Fig.…”

Section: Resultssupporting

confidence: 89%

“…These include the apo form (Apo-FakA_N), the ATP analog bound form (AMP-PNP-FakA_N), and the ADP bound form (ADP-FakA_N). The organization of the FakA N-terminal domain in our structures agrees with recently solved structures of the ligand-bound FakA N-term 29, 30 . Each of these structures formed a similar overall fold comprising a bundle of eight α-helices termed α1: G7-L31, α2: T41-N58, α3: I64-G78, α4: G81-I98, α5: S106-K122, α6: I132-N149, α7: C153-N173 and α9: S186-L202 and a short helix termed α8: A176-V181 (Fig.…”

Section: Resultssupporting

confidence: 89%

“…Indeed, the basic understanding of the composition of the complex and the mechanism by which fatty acid phosphorylation occurs is unknown. Previous structural studies identified key aspects of the FakA and FakB proteins but have not captured how the complex forms the functional unit in response to binding the respective ligands, ATP and FA, and undergoes catalysis 29, 30 .…”

Section: Introductionmentioning

confidence: 99%

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

Myers,

Xu,

Ryan

et al. 2024

Preprint

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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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“…S4). This matches the published C-term FakA structure (PDB 6W6B, 30 ). The FakA C-term structure consists of both β- sheets and α-helices (Fig.…”

Section: Resultssupporting

confidence: 89%

Section: Resultssupporting

confidence: 89%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

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

Myers,

Xu,

Ryan

et al. 2024

Preprint

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“…The amounts of short-chain acids varied significantly between the three different undefined rich medias analyzed here and other laboratories may find different results with these medias depending on the supplier and potentially the lot number. The BCFA requirement for S. aureus is also satisfied in Bkd-deficient strains by the addition of exogenous BCFA which can be incorporated into PG after their activation by fatty acid kinase ( 36 , 37 ). BCFA are not found in sufficient concentrations in complex-rich medias to support the growth of Bkd-deficient strains ( 29 , 38 ), although medias like BHI broth do contain sources of mammalian straight-chain fatty acids.…”

Section: Discussionmentioning

confidence: 99%

A short-chain acyl-CoA synthetase that supports branched-chain fatty acid synthesis in Staphylococcus aureus

Whaley¹,

Frank²,

Rock³

2023

Journal of Biological Chemistry

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“…FakA and Dak2 are orthologues (Bose et al, 2014;Parsons, Broussard, et al, 2014) and a series of separate mutations at five different positions from 14 isolates of both species were found in the 3 different structural domains (characterised by Subramanian et al, 2022) of the fatty acid kinase which were recovered from both species in biofilm and planktonic conditions.…”

Section: Genotyping the Selected Evolved Isolatesmentioning

confidence: 99%

FakA impacts antiseptic susceptibility inStaphylococcus aureusandEnterococcus faecalis

Gaya,

Felgate,

Siani

et al. 2024

Preprint

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Biocides are widely used to control and prevent healthcare-associated infections and understanding how nosocomial pathogens respond to biocidal agents is key to improving infection prevention and control products and practices. An evolution model was used to study how Staphylococcus aureus and Enterococcus faecalis responded after repeated exposure to sub-lethal concentrations of chlorhexidine digluconate (CHX) and octenidine dihydrochloride (OCT) when grown planktonically and as biofilms. Both pathogens were able to adapt to grow at concentrations above the MIC of both biocides with planktonic lineages able to adapt to survive at higher concentrations of both agents than biofilm lineages. Exposure to CHX was linked with lower biofilm biomass production in E. faecalis although biofilm biomass increased for S. aureus isolates after exposure to both agents. Evolved isolates had no major fitness deficit and only low-level changes to susceptibility to antibiotics were observed after biocide exposure. Sequencing of biocide adapted mutants repeatedly identified mutations within fakA encoding a fatty acid kinase in independent lineages of S. aureus after exposure to both biocides in all conditions. Analogous changes were observed within the homologous gene in parallel experiments with Enterococcus faecalis. Further assays to study the mechanistic basis and relationship to phospholipid production showed that evolved isolates with fakA mutations accumulated less ethidium bromide than parent strains, exhibited altered cell envelope morphology and decreased susceptibility to daptomycin. This data shows important pathogens can evolve limited tolerance to two common biocides but that this has collateral impacts on biofilm formation, colony morphology and fitness. FakA appears to play an important role in biocide tolerance.

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Domain architecture and catalysis of the Staphylococcus aureus fatty acid kinase

Cited by 7 publications

References 49 publications

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

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

A short-chain acyl-CoA synthetase that supports branched-chain fatty acid synthesis in Staphylococcus aureus

FakA impacts antiseptic susceptibility inStaphylococcus aureusandEnterococcus faecalis

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