Branched-chain amino acid metabolism controls membrane phospholipid structure in Staphylococcus aureus

Frank, Matthew W.; Whaley, Sarah G.; Rock, Charles O.

doi:10.1016/j.jbc.2021.101255

Cited by 32 publications

(41 citation statements)

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“…The reactions were performed with nonradioactive substrates, the samples were acid precipitated, and the pellets hydrolyzed with Asp-N protease. This process liberates a tripeptide harboring the prosthetic group and attached acyl chains that were then separated and detected by LC-MS/MS ( 17 , 18 , 19 ). Acetyl-ACP was not detected in assays without MadA and was robustly detected in assays containing MadA ( Fig.…”

Section: Resultsmentioning

confidence: 99%

“…The cultures were incubated at 37 °C until they reached an A 600 = 0.6. The extraction method, liquid chromatography column and gradient, mass spectrometry parameters, and the Q1 and Q3 masses for the acyl-CoA were exactly as described ( 17 ).…”

Section: Methodsmentioning

confidence: 99%

“…The samples were prepared for acyl-ACP quantification, as previously described ( 17 , 18 ). One milliliter aliquots of cultures at mid-log phase growth were removed to a 1.5 ml Eppendorf tube containing 250 μl ice cold 10% trichloroacetic acid.…”

Section: Methodsmentioning

confidence: 99%

“…Proteolysis was quenched with 15 μl methanol. The liquid chromatography column and solvent gradient, mass spectrometry parameters, and Q1 and Q3 masses for acyl-ACPs were exactly as described ( 17 ).…”

Section: Methodsmentioning

confidence: 99%

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Malonyl-acyl carrier protein decarboxylase activity promotes fatty acid and cell envelope biosynthesis in Proteobacteria

Whaley¹,

Radka²,

Subramanian³

et al. 2021

Journal of Biological Chemistry

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Bacterial fatty acid synthesis in Escherichia coli is initiated by the condensation of an acetyl-CoA with a malonyl-acyl carrier protein (ACP) by the β-ketoacyl-ACP synthase III enzyme, FabH. E. coli Δ fabH knockout strains are viable because of the yiiD gene that allows FabH-independent fatty acid synthesis initiation. However, the molecular function of the yiiD gene product is not known. Here, we show the yiiD gene product is a m alonyl- A CP d ecarboxylase (MadA). MadA has two independently folded domains: an amino-terminal N -acetyl transferase (GNAT) domain (MadA N ) and a carboxy-terminal hot dog dimerization domain (MadA C ) that encodes the malonyl-ACP decarboxylase function. Members of the proteobacterial Mad protein family are either two domain MadA (GNAT-hot dog) or standalone MadB (hot dog) decarboxylases. Using structure-guided, site-directed mutagenesis of MadB from Shewanella oneidensis , we identified Asn45 on a conserved catalytic loop as critical for decarboxylase activity. We also found that MadA, MadA C , or MadB expression all restored normal cell size and growth rates to an E. coli Δ fabH strain, whereas the expression of MadA N did not. Finally, we verified that GlmU, a bifunctional glucosamine-1-phosphate N -acetyl transferase/ N -acetyl-glucosamine-1-phosphate uridylyltransferase that synthesizes the key intermediate UDP-GlcNAc, is an ACP binding protein. Acetyl-ACP is the preferred glucosamine-1-phosphate N -acetyl transferase/ N -acetyl-glucosamine-1-phosphate uridylyltransferase substrate, in addition to being the substrate for the elongation-condensing enzymes FabB and FabF. Thus, we conclude that the Mad family of malonyl-ACP decarboxylases supplies acetyl-ACP to support the initiation of fatty acid, lipopolysaccharide, peptidoglycan, and enterobacterial common antigen biosynthesis in Proteobacteria.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

Malonyl-acyl carrier protein decarboxylase activity promotes fatty acid and cell envelope biosynthesis in Proteobacteria

Whaley¹,

Radka²,

Subramanian³

et al. 2021

Journal of Biological Chemistry

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“…Metabolomics results also revealed some enriched pathways in glucose and carbon metabolism, such as C5-branched dibasic acid metabolism. Previous studies suggested that multiple factors in amino acid metabolism moonlight as regulators in the expressions of genes regulating bacterial virulence ( Lomas-Lopez et al, 2007 ; Frank et al, 2021 ). Our findings also showed that amino acid metabolism was significantly enriched according to KEGG analysis of metabolome and transcriptome data.…”

Section: Discussionmentioning

confidence: 99%

Insights Into the Impact of Small RNA SprC on the Metabolism and Virulence of Staphylococcus aureus

Zhou

Zhao

et al. 2022

Front. Cell. Infect. Microbiol.

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AimOur previous proteomic analysis showed that small RNA SprC (one of the small pathogenicity island RNAs) of Staphylococcus aureus possesses the ability to regulate the expression of multiple bacterial proteins. In this study, our objective was to further provide insights into the regulatory role of SprC in gene transcription and metabolism of S. aureus.MethodsGene expression profiles were obtained from S. aureus N315 wild-type and its sprC deletion mutant strains by RNA-sequencing (RNA-seq), and differentially expressed genes (DEGs) were screened by R language with a |log2(fold change)| ≥1 and a false discovery rate (FDR) ≤ 0.05. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were carried out to understand the significance of the DEGs. The quality of RNA-seq was further verified by quantitative real-time PCR (qRT-PCR), mRNA target prediction, metabolomics analysis and transcript-level expression analysis of genes of sprC complementation strain.ResultsA total of 2497 transcripts were identified, of which 60 transcripts expressions in sprC knockout strain were significantly different (37 up-regulated and 23 down-regulated DEGs). GO analysis showed that the functions of these DEGs were mainly concentrated in the biological process and molecular function related to metabolism and pathogenesis, and a higher number of genes were involved in the oxidation-reduction process, catalytic activity and binding. KEGG pathways enrichment analysis demonstrated that metabolism and pathogenesis were the most affected pathways, such as metabolic pathways, biosynthesis of secondary metabolites, purine metabolism, fructose and mannose metabolism and S. aureus infection. The qRT-PCR results of the DEGs with defined functions in the sprC deletion and complementation strains were in general agreement with those obtained by RNA-seq. Metabolomics analysis revealed 77 specific pathways involving metabolic pathways. Among them, many, such as metabolic pathways, biosynthesis of secondary metabolites and purine metabolism, were consistent with those enriched in the RNA-seq analysis.ConclusionThis study offered valuable and reliable information about the regulatory roles of SprC in S. aureus biology through transcriptomics and metabolomics analysis. These results may provide clues for new potential targets for anti-virulence adjuvant therapy on S. aureus infection.

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Tribovoltaic Effect Strengthened Microwave Catalytic Antibacterial Composite Hydrogel

Zheng,

Liu,

Mao

et al. 2024

Adv Healthcare Materials

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Microwave (MW) therapy is an emerging therapy with high efficiency and deep penetration to combat the crisis of bacterial resistance. However, as the energy of MW is too low to induce electron transition, the mechanism of MW catalytic effect remains ambiguous. Herein, a cerium‐based metal–organic framework (MOF) is fabricated and used in MW therapy. The MW‐catalytic performance of CeTCPP is largely dependent on the ions in the liquid environment, and the electron transition is achieved through a “tribovoltaic effect” between water molecules and CeTCPP. By this way, CeTCPP can generate reactive oxygen species (ROS) in saline under pulsed MW irradiation, showing 99.9995 ± 0.0002% antibacterial ratio against Staphylococcus aureus (S. aureus) upon two cycles of MW irradiation. Bacterial metabolomics further demonstrates that the diffusion of ROS into bacteria led to the bacterial metabolic disorders. The bacteria are finally killed due to “amino acid starvation”. In order to improve the applicability of CeTCPP, It is incorporated into alginate‐based hydrogel, which maintains good MW catalytic antibacterial efficiency and also good biocompatibility. Therefore, this work provides a comprehensive instruction of using CeTCPP in MW therapy, from mechanism to application. This work also provides new perspectives for the design of antibacterial composite hydrogel.

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Branched-chain amino acid metabolism controls membrane phospholipid structure in Staphylococcus aureus

Cited by 32 publications

References 56 publications

Malonyl-acyl carrier protein decarboxylase activity promotes fatty acid and cell envelope biosynthesis in Proteobacteria

Malonyl-acyl carrier protein decarboxylase activity promotes fatty acid and cell envelope biosynthesis in Proteobacteria

Insights Into the Impact of Small RNA SprC on the Metabolism and Virulence of Staphylococcus aureus

Tribovoltaic Effect Strengthened Microwave Catalytic Antibacterial Composite Hydrogel

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