Permeability of the peroxisomal membrane to cofactors of beta-oxidation. Evidence for the presence of a pore-forming protein.
Peroxisomes were purified from livers of clofibrate-treated rats. Permeability measurements on the isolated organelles revealed that peroxisomes are permeable to small solutes, including sucrose and the cofactors for fatty acid oxidation NAD+, CoA, ATP, and carnitine. The intraperoxisomal distribution volume was equal for all solutes. Peroxisomal solute uptake was rapid, not saturable and not visibly influenced by temperature. NAD+ and carnitine uptake in the solute accessible volume was not diminished by a variety of analogs and inhibitors. Subfractionation of peroxisomes and reconstitution of the subfractions into liposomes preloaded with solutes made the liposomes reconstituted with the integral membrane protein fraction, but not those reconstituted with the other subperoxisomal protein fractions, permeable to the same solutes that entered intact peroxisomes. Solute leakage from the preloaded liposomes was rapid and not visibly influenced by temperature. Leakage activity was destroyed by heat treatment of the integral membrane protein fraction and was not present in lipid extracts of the membrane. Separation of the integral membrane proteins on sucrose density gradients and reconstitution of the gradient fractions into liposomes indicated that the leakage activity was caused by a polypeptide of rather low molecular weight. The gradient distribution of leakage activity corresponded most closely to the presence of a 22- and a 28-kDa polypeptide. Our experiments indicate that the nonspecific permeability of the peroxisomal membrane to small solutes is based on the presence in the membrane of a nonselective pore-forming protein.
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D001425:Bacterial Outer Membrane Proteins / Q000378:metabolism*; D002331:Carnitine / Q000378:metabolism; D002458:Cell Fractionation; D002463:Cell Membrane Permeability; D003065:Coenzyme A / Q000378:metabolism; D007425:Intracellular Membranes / Q000378:metabolism; D008081:Liposomes / Q000378:metabolism; D008099:Liver / Q000648:ultrastructure*; D008297:Male; D008565:Membrane Proteins / Q000302:isolation & purification / Q000378:metabolism; D008830:Microbodies / Q000378:metabolism*; D009243:NAD / Q000378:metabolism; D010084:Oxidation-Reduction; D018272:Porins; D051381:Rats; D013395:Sucrose / Q000378:metabolism
D001425:Bacterial Outer Membrane Proteins; D008081:Liposomes; D008565:Membrane Proteins; D018272:Porins; D009243:NAD; D013395:Sucrose; D000255:Adenosine Triphosphate; D002331:Carnitine; D003065:Coenzyme A