Properties of the mitochondrial peptide-sensitive cationic channel studied in planar bilayers and patches of giant liposomes.
A voltage-dependent cationic channel of large conductance is observed in phospholipid bilayers formed by the tip-dip method from proteoliposomes derived from mitochondrial membranes. It is blocked by peptide M, a 13 residue peptide having the properties of a mitochondrial signal sequence. To verify the reliability of the experimental approach, mitochondrial membranes from bovine adrenal cortex or porin-deficient mutant yeast were either fused to planar bilayers or incorporated in giant liposomes which were studied by patch clamp. Cationic channels were found with both techniques. They had the same conductance levels and voltage-dependence as those which have been described using the tip-dip method. Moreover, they were similarly blocked by peptide M. The voltage-dependence of block duration was analyzed in planar bilayer and tip-dip records. Results strengthen the idea that peptide M might cross the channel. Other mitochondrial channels were observed in planar bilayers and patch clamp of giant liposomes. Because they were never detected in tip-dip records, they are likely to be inactivated at the surface monolayer used to form the bilayer in this type of experiment.
Thieffry
M
M
Laboratoire de Neurobiologie Cellulaire et Moléculaire, CNRS, Gif sur Yvette, France.
Neyton
J
J
Pelleschi
M
M
Fèvre
F
F
Henry
J P
JP
D000302:Adrenal Cortex / Q000502:physiology*; D000818:Animals; D002417:Cattle; D004553:Electric Conductivity; D007473:Ion Channels / Q000187:drug effects / Q000502:physiology*; D008051:Lipid Bilayers; D008081:Liposomes; D008564:Membrane Potentials / Q000187:drug effects; D008928:Mitochondria / Q000502:physiology*; D010713:Phosphatidylcholines; D010714:Phosphatidylethanolamines; D010718:Phosphatidylserines; D011510:Proteolipids / Q000502:physiology; D012441:Saccharomyces cerevisiae / Q000502:physiology; D014357:Trypsin / Q000494:pharmacology
D007473:Ion Channels; D008051:Lipid Bilayers; D008081:Liposomes; D010713:Phosphatidylcholines; D010714:Phosphatidylethanolamines; D010718:Phosphatidylserines; D011510:Proteolipids; C020485:proteoliposomes; C057561:1-palmitoyl-2-oleoylphosphatidylethanolamine; D014357:Trypsin; C028694:1-palmitoyl-2-oleoylphosphatidylcholine