Kinetic evidence for a heart mitochondrial pore activated by Ca2+, inorganic phosphate and oxidative stress. A potential mechanism for mitochondrial dysfunction during cellular Ca2+ overload.
Evidence that the Ca2+-induced permeabilization of mitochondria is attributable to a reversible Ca2+-activated pore [Al Nasser & Crompton (1986) Biochem. J. 239, 19-29] has been further investigated. Permeabilization is induced in a wholly synergistic manner by either Ca2+ plus phosphate or Ca2+ plus tert-butyl hydroperoxide. When permeabilization is complete, extramitochondrial [14C]sucrose equilibrates with the matrix space with a half-time of about 800 ms; [14C]mannitol equilibrates at least threefold faster. Permeabilization is essentially fully reversed on Ca2+ chelation with EGTA, when the half time for [14C]sucrose equilibration is increased 600-1400-fold (to 550-1150 s). A pulsed-flow [14C]solute-entrapment technique has been developed to measure the kinetics of EGTA-induced resealing. The technique incorporates a suitable choice of [14C]solute and an appropriate model for data analysis, and is competent to measure permeation state changes occurring in 100 ms. The data obtained are consistent with exponential resealing of mitochondria in which pores of any single mitochondria close with a high degree of synchrony. The rate of resealing is increased about eight-fold by ADP (half-time approximately 1 s; Km approximately 30 microM). CoA, Mg2+, AMP and also ATP, when account is taken of ADP arising by hydrolysis, are essentially ineffective. It is concluded that heart mitochondria do contain a pore whose permeation state is controlled over an approximate 1000-fold range by Ca2+ and other factors including phosphate, oxidative stress and ADP. The possible involvement of the pore in reoxygenation-induced injury in heart is discussed.
Crompton
M
M
Department of Biochemistry, University College London, England.
Costi
A
A
D000244:Adenosine Diphosphate / Q000494:pharmacology; D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D001711:Biotransformation / Q000187:drug effects; D002118:Calcium / Q000494:pharmacology*; D015220:Calcium Channels / Q000187:drug effects* / Q000378:metabolism; D004533:Egtazic Acid / Q000494:pharmacology; D007425:Intracellular Membranes / Q000378:metabolism; D007700:Kinetics; D008353:Mannitol / Q000378:metabolism; D008929:Mitochondria, Heart / Q000378:metabolism*; D010084:Oxidation-Reduction; D010539:Permeability; D010710:Phosphates / Q000494:pharmacology*; D051381:Rats; D011919:Rats, Inbred Strains; D013395:Sucrose / Q000378:metabolism
D015220:Calcium Channels; D010710:Phosphates; D008353:Mannitol; D004533:Egtazic Acid; D013395:Sucrose; D000244:Adenosine Diphosphate; D002118:Calcium