Solubilization and reconstitution of the mitochondrial peptide-sensitive channel.
In addition to the voltage-dependent anion channel (VDAC), mitochondrial outer membranes contain a cationic channel of large conductance, which is blocked by a mitochondrial addressing peptide (peptide-sensitive channel, PSC). Bovine adrenal cortex mitochondria were solubilized in 1.5% octyl beta-glucoside, and membrane vesicles were reconstituted by slow dilution with a low ionic strength buffer. The reconstituted vesicles contained a functional channel possessing the electrical characteristics of the cationic channel, including its sensitivity to the mitochondrial addressing peptide. Important features of the described protocol are the nature of the detergent, its concentration, and the addition of glycerol during the whole procedure. No solubilization could be observed in the presence of cholate.
Fèvre
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Laboratoire de Neurobiologie Cellulaire et Moléculaire, CNRS UPR 23, Gif sur Yvette, France.
Henry
J P
JP
Thieffry
M
M
D000302:Adrenal Cortex / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D003902:Detergents; D005960:Glucosides; D066298:In Vitro Techniques; D007473:Ion Channels / Q000187:drug effects / Q000378:metabolism*; D008928:Mitochondria / Q000378:metabolism*; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry / Q000494:pharmacology; D012995:Solubility
D003902:Detergents; D005960:Glucosides; D007473:Ion Channels; D010455:Peptides; C093986:peptide sensitive channel; C018619:octyl-beta-D-glucoside