Journal
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
Volume 16, Issue 8, Pages 16710-16727Publisher
MDPI AG
DOI: 10.3390/ijms160816710
Keywords
epicatechin gallate; Staphylococcus aureus; membrane phospholipids; membrane asymmetry; beta-lactam susceptibility; bacterial cell division
Funding
- Biotechnology and Biological Sciences Research Council [BB/1005579/1]
- National Institute for Health Research University College London Hospitals Biomedical Research Centre
- Biotechnology and Biological Sciences Research Council [BB/I005579/1] Funding Source: researchfish
- BBSRC [BB/I005579/1] Funding Source: UKRI
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The polyphenol (-)-epicatechin gallate (ECg) inserts into the cytoplasmic membrane (CM) of methicillin-resistant Staphylococcus aureus (MRSA) and reversibly abrogates resistance to -lactam antibiotics. ECg elicits an increase in MRSA cell size and induces thickened cell walls. As ECg partially delocalizes penicillin-binding protein PBP2 from the septal division site, reduces PBP2 and PBP2a complexation and induces CM remodelling, we examined the impact of ECg membrane intercalation on phospholipid distribution across the CM and determined if ECg affects the equatorial, orthogonal mode of division. The major phospholipids of the staphylococcal CM, lysylphosphatidylglycerol (LPG), phosphatidylglycerol (PG), and cardiolipin (CL), were distributed in highly asymmetric fashion; 95%-97% of LPG was associated with the inner leaflet whereas PG (similar to 90%) and CL (similar to 80%) were found predominantly in the outer leaflet. ECg elicited small, significant changes in LPG distribution. Atomic force microscopy established that ECg-exposed cells divided in similar fashion to control bacteria, with a thickened band of encircling peptidoglycan representing the most recent plane of cell division, less distinct ribs indicative of previous sites of orthogonal division and concentric rings and knobbles representing stages of peptidoglycan remodelling during the cell cycle. Preservation of staphylococcal membrane lipid asymmetry and mode of division in sequential orthogonal planes appear key features of ECg-induced stress.
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