Induction of a non-specific permeability transition in mitochondria from Yarrowia lipolytica and Dipodascus (Endomyces) magnusii yeasts.
In this study we used tightly-coupled mitochondria from Yarrowia lipolytica and Dipodascus (Endomyces) magnusii yeasts, possessing a respiratory chain with the usual three points of energy conservation. High-amplitude swelling and collapse of the membrane potential were used as parameters for demonstrating induction of the mitochondrial permeability transition due to opening of a pore (mPTP). Mitochondria from Y. lipolytica, lacking a natural mitochondrial Ca(2+) uptake pathway, and from D. magnusii, harboring a high-capacitive, regulated mitochondrial Ca(2+) transport system (Bazhenova et al. J Biol Chem 273:4372-4377, 1998a; Bazhenova et al. Biochim Biophys Acta 1371:96-100, 1998b; Deryabina and Zvyagilskaya Biochemistry (Moscow) 65:1352-1356, 2000; Deryabina et al. J Biol Chem 276:47801-47806, 2001) were very resistant to Ca(2+) overload. However, exposure of yeast mitochondria to 50-100 microM Ca(2+) in the presence of the Ca(2+) ionophore ETH129 induced collapse of the membrane potential, possibly due to activation of the fatty acid-dependent Ca(2+)/nH(+)-antiporter, with no classical mPTP induction. The absence of response in yeast mitochondria was not simply due to structural limitations, since large-amplitude swelling occurred in the presence of alamethicin, a hydrophobic, helical peptide, forming voltage-sensitive ion channels in lipid membranes. Ca(2+)- ETH129-induced activation of the Ca(2+)/H(+)-antiport system was inhibited and prevented by bovine serum albumin, and partially by inorganic phosphate and ATP. We subjected yeast mitochondria to other conditions known to induce the permeability transition in animal mitochondria, i.e., Ca(2+) overload (in the presence of ETH129) combined with palmitic acid (Mironova et al. J Bioenerg Biomembr 33:319-331, 2001; Sultan and Sokolove Arch Biochem Biophys 386:37-51, 2001), SH-reagents, carboxyatractyloside (an inhibitor of the ADP/ATP translocator), depletion of intramitochondrial adenine nucleotide pools, deenergization of mitochondria, and shifting to acidic pH values in the presence of high phosphate concentrations. None of the above-mentioned substances or conditions induced a mPTP-like pore. It is thus evident that the permeability transition in yeast mitochondria is not coupled with Ca(2+) uptake and is differently regulated compared to the mPTP of animal mitochondria.
Kovaleva
Mariya V
MV
A.N. Bach Institute of Biochemistry, Russian Academy of Sciences, Moscow, Russia.
Sukhanova
Evgeniya I
EI
Trendeleva
Tatyana A
TA
Zyl'kova
Marina V
MV
Ural'skaya
Ludmila A
LA
Popova
Kristina M
KM
Saris
Nils-Erik L
NE
Zvyagilskaya
Renata A
RA
D000408:Alamethicin / Q000494:pharmacology; D017920:Antiporters / Q000378:metabolism; D002118:Calcium / Q000378:metabolism / Q000494:pharmacology; D027682:Cation Transport Proteins / Q000378:metabolism; D003510:Cyclohexanes / Q000494:pharmacology; D055157:Dipodascus / Q000166:cytology*; D007476:Ionophores / Q000494:pharmacology; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology*; D051336:Mitochondrial Membranes / Q000502:physiology*; D010539:Permeability; D025062:Yarrowia / Q000166:cytology*
D017920:Antiporters; D027682:Cation Transport Proteins; D003510:Cyclohexanes; D007476:Ionophores; C027614:calcium-hydrogen antiporters; D000408:Alamethicin; C077946:N,N,N',N'-tetracyclohexyl-3-oxapentanediamide; D002118:Calcium