Dataset Information

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.

SUBMITTER: Niu W

PROVIDER: S-EPMC10227063 | biostudies-literature | 2023 May

REPOSITORIES: biostudies-literature

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

Niu Wei W Zhou Wenchang W Lu Shuo S Vu Trung T Jayaraman Vasanthi V Faraldo-Gómez José D JD Zheng Lei L

Communications biology 20230529 1

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-t ...[more]

PMID: 37248347

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Project description:Mitochondrial Ca2+ handling is accomplished by balancing Ca2+ uptake, primarily via the Ru360-sensitive mitochondrial calcium uniporter (MCU), Ca2+ buffering in the matrix and Ca2+ efflux mainly via Ca2+ ion exchangers, such as the Na+/Ca2+ exchanger (NCLX) and the Ca2+/H+ exchanger (CHE). The mechanism of CHE in cardiac mitochondria is not well-understood and its contribution to matrix Ca2+ regulation is thought to be negligible, despite higher expression of the putative CHE protein, LETM1, compared to hepatic mitochondria. In this study, Ca2+ efflux via the CHE was investigated in isolated rat cardiac mitochondria and permeabilized H9c2 cells. Mitochondria were exposed to (a) increasing matrix Ca2+ load via repetitive application of a finite CaCl2 bolus to the external medium and (b) change in the pH gradient across the inner mitochondrial membrane (IMM). Ca2+ efflux at different matrix Ca2+ loads was revealed by inhibiting Ca2+ uptake or reuptake with Ru360 after increasing number of CaCl2 boluses. In Na+-free experimental buffer and with Ca2+ uptake inhibited, the rate of Ca2+ efflux and steady-state free matrix Ca2+ [mCa2+]ss increased as the number of administered CaCl2 boluses increased. ADP and cyclosporine A (CsA), which are known to increase Ca2+ buffering while maintaining a constant [mCa2+]ss, decreased the rate of Ca2+ efflux via the CHE, with a significantly greater decrease in the presence of ADP. ADP also increased Ca2+ buffering rate and decreased [mCa2+]ss. A change in the pH of the external medium to a more acidic value from 7.15 to 6.8∼6.9 caused a twofold increase in the Ca2+ efflux rate, while an alkaline change in pH from 7.15 to 7.4∼7.5 did not change the Ca2+ efflux rate. In addition, CHE activation was associated with membrane depolarization. Targeted transient knockdown of LETM1 in permeabilized H9c2 cells modulated Ca2+ efflux. The results indicate that Ca2+ efflux via the CHE in cardiac mitochondria is modulated by acidic buffer pH and by total matrix Ca2+. A mechanism is proposed whereby activation of CHE is sensitive to changes in both the matrix Ca2+ buffering system and the matrix free Ca2+ concentration.

Dataset Information

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

Publications

Ca<sup>2+</sup> efflux facilitated by co-transport of inorganic phosphate anion in the H<sup>+</sup>/Ca<sup>2+</sup> antiporter YfkE.

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