Ca2+ efflux facilitated by co-transport of inorganic phosphate anion in the H+/Ca2+ antiporter YfkE

Abstract

Ca²⁺ is an important signaling messenger. In microorganisms, fungi, and plants, H⁺/Ca²⁺ antiporters (CAX) are known to play key roles in the homeostasis of intracellular Ca²⁺ by catalyzing its efflux across the cell membrane. Here, we reveal that the bacterial CAX homolog YfkE transports Ca²⁺ in two distinct modes: a low-flux H⁺/Ca²⁺ exchange mode and a high-flux mode in which Ca²⁺ and phosphate ions are co-transported (1:1) in exchange for H⁺. Coupling with phosphate greatly accelerates the Ca²⁺ efflux activity of YfkE. Our studies reveal that Ca²⁺ and phosphate bind to adjacent sites in a central translocation pathway and lead to mechanistic insights that explain how this CAX alters its conserved alpha-repeat motifs to adopt phosphate as a specific "transport chaperon" for Ca²⁺ translocation. This finding uncovers a co-transport mechanism within the CAX family that indicates this class of proteins contributes to the cellular homeostasis of both Ca²⁺ and phosphate.