The use of proteinases to determine the topological location of cytochrome P-450 in vesicles derived from smooth endoplasmic reticulum of rat liver.
1. The phospholipid bilayer of intact vesicles from smooth endoplasmic reticulum is impermeable to macromolecules. Specific and non-specific proteinases were used to investigate the site of membrane proteins in the transverse plane of the bilayer. 2. When two proteinases were used in conjunction, denaturing effects additional to proteolysis were observed on cytochrome P-450 content and glucose 6-phosphatase activity which did not depend on the integrity of the phospholipid bilayer. 3. When lipid peroxidation was inhibited, these effects were not observed.
D000818:Animals; D002084:Butylated Hydroxytoluene / Q000494:pharmacology; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D010450:Endopeptidases / Q000494:pharmacology*; D004721:Endoplasmic Reticulum / Q000187:drug effects / Q000201:enzymology*; D005296:Ferrous Compounds / Q000494:pharmacology; D005952:Glucose-6-Phosphatase / Q000378:metabolism; D066298:In Vitro Techniques; D008051:Lipid Bilayers / Q000378:metabolism; D008099:Liver / Q000187:drug effects / Q000201:enzymology*; D008297:Male; D008315:Malondialdehyde / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains
D005296:Ferrous Compounds; D008051:Lipid Bilayers; D008565:Membrane Proteins; D002084:Butylated Hydroxytoluene; D008315:Malondialdehyde; D003577:Cytochrome P-450 Enzyme System; D005952:Glucose-6-Phosphatase; D010450:Endopeptidases