Cytochrome P-450 and NADPH-cytochrome P-450 reductase are degraded in the autolysosomes in rat liver.
We have investigated the degradation in rat liver of two typical endoplasmic reticulum (ER) membrane proteins, phenobarbital (PB)-inducible cytochrome P-450 (P-450[PB]) and NADPH-cytochrome P-450 reductase (FP2). Autolysosomes, almost completely free from contamination by the other organelles such as ER, were prepared from leupeptin-treated rat livers according to the method of Furuno et al. (Furuno, K., T. Ishikawa, and K. Kato, 1982, J. Biochem., 91:1943-1950). Quantitative immunoblot analysis showed that these two proteins were found in large amounts in the autolysosomes regardless of PB treatment. The specific content of P-450 (PB) in the autolysosomes changed along with that in the microsomes during and after PB treatment, whereas hardly any P-450(PB) was detected in the cytosol fraction throughout the experiment. We also found a marked increase in the autolysosomal proteins 3 d after cessation of PB treatment when microsomal proteins are degraded most rapidly. Ferritin immunoelectron microscopy revealed directly that when the limiting membranes of the premature autolysosomes were partially broken the smooth vesicles segregated within the autolysosomes were heavily stained with ferritin anti-P-450(PB) conjugates. Thus, for the first time, we could present convincing evidence that P-450(PB) and FP2 are segregated to be degraded in the autolysosomes.
D000818:Animals; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D004790:Enzyme Induction; D007700:Kinetics; D008099:Liver / Q000187:drug effects / Q000378:metabolism*; D008247:Lysosomes / Q000378:metabolism* / Q000648:ultrastructure; D008297:Male; D008854:Microscopy, Electron; D009251:NADPH-Ferrihemoprotein Reductase / Q000378:metabolism*; D010634:Phenobarbital / Q000494:pharmacology; D051381:Rats; D011919:Rats, Inbred Strains
D003577:Cytochrome P-450 Enzyme System; D009251:NADPH-Ferrihemoprotein Reductase; D010634:Phenobarbital