title: Interaction of non-classical detergents with the mitochondrial porin. A new purification procedure and characterization of the pore-forming unit.
abstract: The effect of different families of detergents on the solubilization and purification of the pore-forming protein (porin) of the mitochondrial outer membrane of bovine heart was investigated in detail. With Tritons, dimethylamine oxides and zwittergents, porin solubilization with respect to total mitochondrial membrane protein was more efficient with the more hydrophobic members of each series. With most detergents the protein eluted as protein-detergent micelles in the void volume of hydroxyapatite/celite columns. In contrast, the protein was bound to the column material and was eluted after the addition of salt to the elution buffer when the detergents octylglucoside, zwittergent Z-314 and lauryl(dimethyl)-amine oxide were used. The protein purified in the presence of the latter detergent had a higher pore-forming activity in lipid bilayer membranes compared to porin isolated in the presence of Triton X-100. The binding of porin to the hydroxyapatite/celite column was used to study the lipid content of the active pore-forming complex. The analysis revealed that the complex contained no phospholipid but rather five molecules of cholesterol/polypeptide chain.
authors.affiliation: Department of Pharmaco-Biology, University of Bari, Italy.
authors.forename: V
authors.initials: V
authors.lastname: De Pinto
authors.forename: R
authors.initials: R
authors.lastname: Benz
authors.forename: F
authors.initials: F
authors.lastname: Palmieri
mesh_terms: D000818:Animals; D001425:Bacterial Outer Membrane Proteins / Q000302:isolation & purification*; D002417:Cattle; D002462:Cell Membrane / Q000378:metabolism; D003902:Detergents; D008055:Lipids / Q000302:isolation & purification; D008929:Mitochondria, Heart / Q000378:metabolism*; D018272:Porins; D012995:Solubility; D013501:Surface-Active Agents
chemical_list: D001425:Bacterial Outer Membrane Proteins; D003902:Detergents; D008055:Lipids; D018272:Porins; D013501:Surface-Active Agents