title: Dual Effect of Phosphate Transport on Mitochondrial Ca2+ Dynamics.
abstract: The large inner membrane electrochemical driving force and restricted volume of the matrix confer unique constraints on mitochondrial ion transport. Cation uptake along with anion and water movement induces swelling if not compensated by other processes. For mitochondrial Ca(2+) uptake, these include activation of countertransporters (Na(+)/Ca(2+) exchanger and Na(+)/H(+) exchanger) coupled to the proton gradient, ultimately maintained by the proton pumps of the respiratory chain, and Ca(2+) binding to matrix buffers. Inorganic phosphate (Pi) is known to affect both the Ca(2+) uptake rate and the buffering reaction, but the role of anion transport in determining mitochondrial Ca(2+) dynamics is poorly understood. Here we simultaneously monitor extra- and intra-mitochondrial Ca(2+) and mitochondrial membrane potential (ΔΨm) to examine the effects of anion transport on mitochondrial Ca(2+) flux and buffering in Pi-depleted guinea pig cardiac mitochondria. Mitochondrial Ca(2+) uptake proceeded slowly in the absence of Pi but matrix free Ca(2+) ([Ca(2+)]mito) still rose to ~50 μm. Pi (0.001-1 mm) accelerated Ca(2+) uptake but decreased [Ca(2+)]mito by almost 50% while restoring ΔΨm. Pi-dependent effects on Ca(2+) were blocked by inhibiting the phosphate carrier. Mitochondrial Ca(2+) uptake rate was also increased by vanadate (Vi), acetate, ATP, or a non-hydrolyzable ATP analog (AMP-PNP), with differential effects on matrix Ca(2+) buffering and ΔΨm recovery. Interestingly, ATP or AMP-PNP prevented the effects of Pi on Ca(2+) uptake. The results show that anion transport imposes an upper limit on mitochondrial Ca(2+) uptake and modifies the [Ca(2+)]mito response in a complex manner.
authors.affiliation: From the Division of Cardiology, Department of Medicine, The Johns Hopkins University, Baltimore, Maryland 21205.
authors.forename: An-Chi
authors.initials: AC
authors.lastname: Wei
authors.affiliation: From the Division of Cardiology, Department of Medicine, The Johns Hopkins University, Baltimore, Maryland 21205.
authors.forename: Ting
authors.initials: T
authors.lastname: Liu
authors.affiliation: From the Division of Cardiology, Department of Medicine, The Johns Hopkins University, Baltimore, Maryland 21205 bor@jhmi.edu.
authors.forename: Brian
authors.initials: B
authors.lastname: O'Rourke
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002118:Calcium / Q000378:metabolism*; D006168:Guinea Pigs; D017136:Ion Transport; D008929:Mitochondria, Heart / Q000378:metabolism*; D063154:Mitochondrial Dynamics; D010710:Phosphates / Q000378:metabolism*
chemical_list: D010710:Phosphates; D000255:Adenosine Triphosphate; D002118:Calcium
keywords: ATP; bioenergetics; calcium; calcium buffer; calcium uniporter; metabolism; mitochondrial transport; phosphate carrier