Cd(2+) extrusion by P-type Cd(2+)-ATPase of Staphylococcus aureus 17810R via energy-dependent Cd(2+)/H(+) exchange...

Cd(2+) is highly toxic to Staphylococcus aureus since it blocks dithiols in cytoplasmic 2-oxoglutarate dehydrogenase complex (ODHC) participating in energy conservation process. However, S. aureus 17810R is Cd(2+)-resistant due to possession of cadA-coded Cd(2+) efflux system, recognized here as P-type Cd(2+)-ATPase. This Cd(2+) pump utilizing cellular energy-ATP, ∆μ H (+) (electrochemical proton potential) and respiratory protons, extrudes Cd(2+) from cytoplasm to protect dithiols in ODHC, but the mechanism of Cd(2+) extrusion remains unknown. Here we propose that two Cd(2+) taken up by strain 17810R via Mn(2+) uniporter down membrane potential (∆ψ) generated during glutamate oxidation in 100 mM phosphate buffer (high PiB) are trapped probably by high affinity sites in cytoplasmic domain of Cd(2+)-ATPase, forming SCdS. This stops Cd(2+) transport towards dithiols in ODHC, allowing undisturbed NADH production, its oxidation and energy conservation, while ATP could change orientation of SCdS towards facing transmembrane channel. Now, increased number of Pi-dependent protons pumped electrogenically via respiratory chain and countertransported through the channel down ∆ψ, extrude two trapped cytoplasmic Cd(2+), which move to low affinity sites, being then extruded into extracellular space via ∆ψ-dependent Cd(2+)/H(+) exchange. In 1 mM phosphate buffer (low PiB), external Cd(2+) competing with decreased number of Pi-dependent protons, binds to ψs of Cd(2+)-ATPase channel, enters cytoplasm through the channel down ∆ψ via Cd(2+)/Cd(2+) exchange and blocks dithiols in ODHC. However, Mg(2+) pretreatment preventing external Cd(2+) countertransport through the channel down ∆ψ, allowed undisturbed NADH production, its oxidation and extrusion of two cytoplasmic Cd(2+) via Cd(2+)/H(+) exchange, despite low PiB.