On the involvement of cytochrome P-450 in the binding of ribosomes to a subfraction of rat-liver rapidly sedimenting endoplasmic reticulum.
Rat liver endoplasmic reticulum has been separated into four ribosome-containing subfractions, two from rapidly sedimentation endoplasmic reticulum and two from the microsomes, by differential centrifugation and sucrose density centrifugation. Ribosomes from one of the rapidly sedimenting subfractions were extracted by Trion X-100 as a complex with cytochrome P-450, optimally at a detergent protein ratio of 2/1 (w/w). Upon extraction approximately 50% of the cytochrome P-450 in the membrane appeared complex-bound to ribosomes, and, maximally, 6-7 subunit molecules of the cytochrome were attached per ribosome. The specific concentration of cytochrome P-450 on these ribosomes was 2.5-times higher than in the parent membrane. Cytochrome b5, glucose-6-phosphatase, NADPH-cytochrome c reductase, NADH-ferricyanide reductase, cytochrome oxidase and phospholipids were present in small or trace amounts on the ribosomes in relation to cytochrome P-450. Ribosomes extracted from other subfractions contained much less bound cytochrome P-450. Phenobarbital treatment induced an increase in the cytochrome P-450 content that was different for the various subfractions. This increase could not be correlated with changes in the amounts of cytochrome-ribosome complexes released by detergent. We propose that cytochrome P-450 is part of a specific binding site in the membrane for a fraction of the ribosomes attached to the endoplasmic reticulum. The ribosomes may be anchored to cytochrome P-450 via nascent chain proteins.
D000818:Animals; D001665:Binding Sites; D002458:Cell Fractionation; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D004721:Endoplasmic Reticulum / Q000187:drug effects / Q000378:metabolism*; D008099:Liver / Q000378:metabolism*; D008297:Male; D008862:Microsomes, Liver / Q000187:drug effects / Q000378:metabolism; D010634:Phenobarbital / Q000494:pharmacology; D011092:Polyethylene Glycols; D051381:Rats; D012270:Ribosomes / Q000187:drug effects / Q000378:metabolism*
D011092:Polyethylene Glycols; D003577:Cytochrome P-450 Enzyme System; D010634:Phenobarbital