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- W2912150777 abstract "Streptococcus sanguinis is an opportunistic bacterium living beneficially in the human mouth but forming heart valve vegetations when in the blood stream, leading to infective endocarditis. It has been suggested that Mn and Fe are cofactors in the enfeeblement of this bacterium. To delineate the abundance of available metal ions relative to the intracellular protein content, we developed a protocol involving two spectroscopic techniques: Inductively Coupled Plasma-Optical Emission Spectrometry (ICP-OES) and Electron Paramagnetic Resonance (EPR) spectroscopy. Our strategy facilitates direct measurement of intact whole cell samples for metal speciation. ICP-OES enables quantification of the total metal concentration, whereas EPR allows for differentiation and characterization of Fe and Mn oxidation states. Addition of the cell-permeable biological siderophore, deferoxamine, shifts the metal oxidation state equilibrium (from reduced to oxidized) to aid in the quantification of intracellular metal pools. Through this methodology, the influence of exogenous metals on intracellular metal speciation and oxidative maintenance in S. sanguinis can be determined. Previously, it was demonstrated that the SsaB lipoprotein facilitates intracellular Mn accumulation, which can be correlated with aerobic S. sanguinis virulence. We have now established that SsaACB is a Mn/Fe transport system. We have prepared an ATP-binding cassette transporter knockout (ΔssaACB) in order to clarify the mechanism of Mn/Fe import and the effect of metal ion availability on the intracellular redox environment. Our research indicates that SsaACB is responsible for both Fe and Mn transport, and that Mn is pertinent for oxidative maintenance, highlighted through complementary studies conducted under anaerobic conditions. Advanced studies analyzing SsaB structural changes and competitive metal binding are also under investigation to elucidate the role of SsaB and metals in the virulence of S. sanguinis which will provide insight into the health of the oral cavity." @default.
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- W2912150777 date "2019-02-01" @default.
- W2912150777 modified "2023-09-27" @default.
- W2912150777 title "Intracellular Metal Speciation in Streptococcus sanguinis is Pivotal for Redox Maintenance" @default.
- W2912150777 doi "https://doi.org/10.1016/j.bpj.2018.11.890" @default.
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