Insertion of epicatechin gallate into the cytoplasmic membrane of methicillin-resistant Staphylococcus aureus disrupts penicillin-binding protein (PBP) 2a-mediated beta-lactam resistance by delocalizing PBP2.
Epicatechin gallate (ECg) sensitizes methicillin-resistant Staphylococcus aureus (MRSA) to oxacillin and other beta-lactam agents; it also reduces the secretion of virulence-associated proteins, prevents biofilm formation, and induces gross morphological changes in MRSA cells without compromising the growth rate. MRSA is resistant to oxacillin because of the presence of penicillin-binding protein 2a (PBP2a), which allows peptidoglycan synthesis to continue after oxacillin-mediated acylation of native PBPs. We show that ECg binds predominantly to the cytoplasmic membrane (CM), initially decreasing the fluidity of the bilayer, and induces changes in gene expression indicative of an attempt to preserve and repair a compromised cell wall. On further incubation, the CM is reorganized; the amount of lysylphosphatidylglycerol is markedly reduced, with a concomitant increase in phosphatidylglycerol, and the proportion of branched chain fatty acids increases, resulting in a more fluid structure. We found no evidence that ECg modulates the enzymatic activity of PBP2a through direct binding to the protein but determined that PBP2 is delocalized from the FtsZ-anchored cell wall biosynthetic machinery at the septal division site following intercalation into the CM. We argue that many features of the ECg-induced phenotype can be explained by changes in the fluid dynamics of the CM.
School of Pharmacy, University of London, London WC1N 1AX, United Kingdom.
Patricia
P
Bernal
Sandrine
S
Lemaire
Mariana G
MG
Pinho
Shahriar
S
Mobashery
Jason
J
Hinds
Peter W
PW
Taylor
D016880:Anisotropy; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D002392:Catechin / Q000031:analogs & derivatives* / Q000737:chemistry; D002473:Cell Wall / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism*; D024881:Drug Resistance, Bacterial; D015964:Gene Expression Regulation, Bacterial; D008239:Lysine / Q000737:chemistry; D008712:Methicillin / Q000494:pharmacology*; D055624:Methicillin-Resistant Staphylococcus aureus / Q000378:metabolism*; D008856:Microscopy, Fluorescence / Q000379:methods; D020411:Oligonucleotide Array Sequence Analysis; D046915:Penicillin-Binding Proteins / Q000737:chemistry*; D010715:Phosphatidylglycerols / Q000737:chemistry; D047090:beta-Lactams / Q000378:metabolism*
D000900:Anti-Bacterial Agents; D046915:Penicillin-Binding Proteins; D010715:Phosphatidylglycerols; D047090:beta-Lactams; C002285:lysylphosphatidylglycerol; D002392:Catechin; C062669:epicatechin gallate; D008239:Lysine; D008712:Methicillin