Permissive Fatty Acid Incorporation Promotes Staphylococcal Adaptation to FASII Antibiotics in Host Environments

Kénanian, Gérald; Morvan, Claire; Weckel, Antonin; Pathania, Amit; Anba‐Mondoloni, Jamila; Halpern, David; Gaillard, Marine; Solgadi, Audrey; Dupont, Laëtitia; Henry, Céline; Poyart, Claire; Fouet, Agnès; Lamberet, Gilles; Gloux, Karine; Gruss, Alexandra

doi:10.1016/j.celrep.2019.11.071

Cited by 38 publications

(113 citation statements)

References 57 publications

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“…Thesuitability of saFabI inhibitors for treating septicemic S. aureus infections in the clinic has been debated. [29] However,k alimantacin Ah as been shown to be effective against nasal S. aureus infections and therefore holds promise for the treatment of skin and soft tissue infections. [15] Our findings may thus offer inspiration for the design and development of an ew generation of saFabI inhibitors that can be used to combat the escalating public health crisis caused by multidrug-resistant S. aureus infections.…”

Section: Resultsmentioning

confidence: 99%

The Kalimantacin Polyketide Antibiotics Inhibit Fatty Acid Biosynthesis in Staphylococcus aureus by Targeting the Enoyl‐Acyl Carrier Protein Binding Site of FabI

et al. 2020

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The enoyl‐acyl carrier protein reductase enzyme FabI is essential for fatty acid biosynthesis in Staphylococcus aureus and represents a promising target for the development of novel, urgently needed anti‐staphylococcal agents. Here, we elucidate the mode of action of the kalimantacin antibiotics, a novel class of FabI inhibitors with clinically‐relevant activity against multidrug‐resistant S. aureus. By combining X‐ray crystallography with molecular dynamics simulations, in vitro kinetic studies and chemical derivatization experiments, we characterize the interaction between the antibiotics and their target, and we demonstrate that the kalimantacins bind in a unique conformation that differs significantly from the binding mode of other known FabI inhibitors. We also investigate mechanisms of acquired resistance in S. aureus and identify key residues in FabI that stabilize the binding of the antibiotics. Our findings provide intriguing insights into the mode of action of a novel class of FabI inhibitors that will inspire future anti‐staphylococcal drug development.

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Section: Resultsmentioning

confidence: 99%

The Kalimantacin Polyketide Antibiotics Inhibit Fatty Acid Biosynthesis in Staphylococcus aureus by Targeting the Enoyl‐Acyl Carrier Protein Binding Site of FabI

et al. 2020

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“…Although our focus here is on the effect of serum lipids on the S. aureus cell surface, we have no doubt that serum has major impacts on the physiology of S. aureus. For example, Kénanian et al showed that when S. aureus was fed a mixture of exogenous fatty acids, bacterial membrane integrity was perturbed, and stress resulted (38). Supplementation of medium with serum offset the detrimental effects induced by exogenous fatty acids and improved fitness.…”

Section: Discussionmentioning

confidence: 99%

Lipidomic and Ultrastructural Characterization of the Cell Envelope of Staphylococcus aureus Grown in the Presence of Human Serum

Hines

Alvarado

Chen

et al. 2020

mSphere

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Staphylococcus aureus can incorporate exogenous straight-chain unsaturated and saturated fatty acids (SCUFAs and SCFAs, respectively) to replace some of the normally biosynthesized branched-chain fatty acids and SCFAs. In this study, the impact of human serum on the S. aureus lipidome and cell envelope structure was comprehensively characterized. When S. aureus was grown in the presence of 20% human serum, typical human serum lipids, such as cholesterol, sphingomyelin, phosphatidylethanolamines, and phosphatidylcholines, were present in the total lipid extracts. Mass spectrometry showed that SCUFAs were incorporated into all major S. aureus lipid classes, i.e., phosphatidylglycerols, lysyl-phosphatidylglycerols, cardiolipins, and diglucosyldiacylglycerols. Heat-killed S. aureus retained fewer serum lipids and failed to incorporate SCUFAs, suggesting that association and incorporation of serum lipids with S. aureus require a living or nondenatured cell. Cytoplasmic membranes isolated from lysostaphin-produced protoplasts of serum-grown cells retained serum lipids, but washing cells with Triton X-100 removed most of them. Furthermore, electron microscopy studies showed that serum-grown cells had thicker cell envelopes and associated material on the surface, which was partially removed by Triton X-100 washing. To investigate which serum lipids were preferentially hydrolyzed by S. aureus lipases for incorporation, we incubated individual serum lipid classes with S. aureus and found that cholesteryl esters (CEs) and triglycerides (TGs) are the major donors of the incorporated fatty acids. Further experiments using purified Geh lipase confirmed that CEs and TGs were the substrates of this enzyme. Thus, growth in the presence of serum altered the nature of the cell surface with implications for interactions with the host. IMPORTANCE Comprehensive lipidomics of S. aureus grown in the presence of human serum suggests that human serum lipids can associate with the cell envelope without being truly integrated into the lipid membrane. However, fatty acids derived from human serum lipids, including unsaturated fatty acids, can be incorporated into lipid classes that can be biosynthesized by S. aureus itself. Cholesteryl esters and triglycerides are found to be the major source of incorporated fatty acids upon hydrolysis by lipases. These findings have significant implications for the nature of the S. aureus cell surface when grown in vivo. Changes in phospholipid and glycolipid abundances and fatty acid composition could affect membrane biophysics and function and the activity of membrane-targeting antimicrobials. Finally, the association of serum lipids with the cell envelope has implications for the physicochemical nature of the cell surface and its interaction with host defense systems.

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“…Gruss and co-workers found that addition of exogenous fatty acids promotes resistance to FASII antibiotics by S. aureus and selection of resistant strains that bypass FASII inhibition (12, 32). The same group showed that exogenous fatty acids could occupy both the sn -1 and sn -2 positions of PG when cells were grown in Brain Heart Infusion broth supplemented with C14:0, C16:0, and C18:1, or serum (33). This seemingly disproves the essentiality of the requirement for biosynthesized fatty acid anteiso C15:0 at the sn -2 position (15, 34) and undermines the viability of inhibitors of the FASII pathway as useful therapeutic agents (33).…”

Section: Discussionmentioning

confidence: 99%

“…The same group showed that exogenous fatty acids could occupy both the sn -1 and sn -2 positions of PG when cells were grown in Brain Heart Infusion broth supplemented with C14:0, C16:0, and C18:1, or serum (33). This seemingly disproves the essentiality of the requirement for biosynthesized fatty acid anteiso C15:0 at the sn -2 position (15, 34) and undermines the viability of inhibitors of the FASII pathway as useful therapeutic agents (33). In this work, we also observed lipid species containing no C15:0, such as PG 32:1 (18:1/14:0), PG 34:1 (18:1/16:0 and 20:1/14:0), and PG 36:1 (18:1/18:0, 20:1/16:0, and 22:1/14:0), which supports the notion that anteiso C15:0 is not essential.…”

Section: Discussionmentioning

confidence: 99%

Lipidomic and Ultrastructural Characterization of Cell Envelope ofStaphylococcus aureusGrown in the Presence of Human Serum

Hines

Alvarado

Chen

et al. 2020

Preprint

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26Staphylococcus aureus can incorporate exogenous straight-chain unsaturated and 27 saturated fatty acids (SCUFAs and SCFAs, respectively) to replace some of the normally 28 biosynthesized branched-chain fatty acids and SCFAs. In this study, the impact of human serum 29 on the S. aureus lipidome and cell envelope structure was comprehensively characterized. When 30 grown in the presence of 20% human serum, typical human serum lipids, such as cholesterol, 31 sphingomyelin, phosphatidylethanolamines, and phosphatidylcholines, were present in the total 32 lipid extracts. Mass spectrometry showed that SCUFAs were incorporated into all major S. 33 aureus lipid classes, i.e., phosphatidylglycerols, lysyl-phosphatidylglycerols, cardiolipins, and 34 diglucosyldiacylglycerols. Heat-killed S. aureus retained much fewer serum lipids and failed to 35 incorporate SCUFAs, suggesting that association and incorporation of serum lipids with S. 36 aureus requires a living or non-denatured cell. Cytoplasmic membranes isolated from 37 lysostaphin-produced protoplasts of serum-grown cells retained serum lipids, but washing cells 38 with Triton X-100 removed most of them. Furthermore, electron microscopy studies showed that 39 serum-grown cells had thicker cell envelopes and associated material on the surface, which was 40 partially removed by Triton X-100 washing. To investigate which serum lipids were 41 preferentially hydrolyzed by S. aureus lipases for incorporation, we incubated individual serum 42 lipid classes with S. aureus and found that cholesteryl esters (CEs) and triglycerides (TGs) are 43 the major donors of the incorporated fatty acids. Further experiments using purified Geh lipase 44 confirmed CEs and TGs being the substrates of this enzyme. Thus, growth in the presence of 45 serum altered the nature of the cell surface with implications for interactions with the host. 46 Page 3 of 38 IMPORTANCE 47Comprehensive lipidomics of S. aureus grown in the presence of human serum suggests 48 human serum lipids can associate with the cell envelope without being truly integrated into the 49 lipid membrane. However, fatty acids-derived from human serum lipids, including unsaturated 50 fatty acids, can be incorporated into lipid classes that can be biosynthesized by S. aureus itself. 51Cholesteryl esters and triglycerides are found to be the major source of incorporated fatty acids 52 upon hydrolysis by lipases. These findings have significant implications for the nature of the S. 53 aureus cell surface when grown in vivo. Changes in phospholipid and glycolipid abundances and 54 fatty acid composition could affect membrane biophysics and function and the activity of 55 membrane-targeting antimicrobials. Finally, the association of serum lipids with the cell envelope 56 has implications for the physicochemical nature of the cell surface and its interaction with host 57 defense systems. 58 KEYWORDS 59 Lipidomics, human serum lipids, fatty acid incorporation, lipid association, cell envelope 60 structure 61 62 63 64 65 6...

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Permissive Fatty Acid Incorporation Promotes Staphylococcal Adaptation to FASII Antibiotics in Host Environments

Cited by 38 publications

References 57 publications

The Kalimantacin Polyketide Antibiotics Inhibit Fatty Acid Biosynthesis in Staphylococcus aureus by Targeting the Enoyl‐Acyl Carrier Protein Binding Site of FabI

The Kalimantacin Polyketide Antibiotics Inhibit Fatty Acid Biosynthesis in Staphylococcus aureus by Targeting the Enoyl‐Acyl Carrier Protein Binding Site of FabI

Lipidomic and Ultrastructural Characterization of the Cell Envelope of Staphylococcus aureus Grown in the Presence of Human Serum

Lipidomic and Ultrastructural Characterization of Cell Envelope ofStaphylococcus aureusGrown in the Presence of Human Serum

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