Blockade of a mitochondrial cationic channel by an addressing peptide: an electrophysiological study.
A voltage-dependent cationic channel of large conductance is observed in phospholipid bilayers formed at the tip of microelectrodes from proteoliposomes derived from mitochondrial membranes. This channel was blocked by a 13-residue peptide with the sequence of the amino terminal extremity of the nuclear-coded subunit IV of cytochrome c oxidase. The blockade was reversible, voltage- and dose-dependent. The peptide did not affect the activity of a Torpedo chloride channel observed under the same conditions. From experiments with phospholipid monolayers, it is unlikely that the peptide inserts into bilayers under the experimental conditions used. The blockade was observed from both sides of the membrane, being characterized by more frequent transitions to the lower conductance states, and a maximum effect was observed around 0 mV. Channels, the gating mechanism of which had been eliminated by exposure to trypsin, were also blocked by the peptide. For trypsinized channels, the duration of the closure decreased and the blockade saturated at potentials below -30 mV. These observations are consistent with a translocation of the peptide through the channel. Dynorphin B, which has the same length and charge as the peptide, had some blocking activity. Introduction of negative charges in the peptide by succinylation suppressed the activity.
Henry
J P
JP
Laboratoire de Neurobiologie Physico-Chimique, Institut de Biologie Physico-Chimique, Paris, France.
Chich
J F
JF
Goldschmidt
D
D
Thieffry
M
M
D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D018118:Chloride Channels; D002712:Chlorides / Q000378:metabolism*; D004558:Electric Stimulation; D004594:Electrophysiology; D005136:Eye Proteins / Q000378:metabolism*; D066298:In Vitro Techniques; D015640:Ion Channel Gating / Q000187:drug effects*; D007473:Ion Channels / Q000187:drug effects / Q000378:metabolism; D007672:Kidney Cortex / Q000187:drug effects / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism*; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism*; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000378:metabolism*; D014101:Torpedo
D018118:Chloride Channels; D002712:Chlorides; D005136:Eye Proteins; D007473:Ion Channels; D008565:Membrane Proteins; D010446:Peptide Fragments; C052615:peptide M, retinal S antigen