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Inhibition of KdpFABC by KdpD. KdpFABC constructs were purified from E. coli LB2003 cells, which natively express kdpDE , or from E. coli TK2281 cells, in which the kdpFABCDE operon is deleted. In the latter, KdpD variants were reintroduced as indicated by expression from an orthogonal plasmid. Normalized ATPase activity of purified KdpFABC constructs relative to the activity of WT KdpFABC extracted from E. coli LB2003 cells. Bars indicate the mean and error bars denote the standard deviation from biological triplicates ( n = 3).

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