Characterization of the integral membrane polypeptides of rat liver peroxisomes isolated from untreated and clofibrate-treated rats.
We have characterized the integral membrane polypeptides of liver peroxisomes from untreated rats and rats treated with clofibrate, a peroxisome proliferator. Membranes, prepared by treatment of purified peroxisomes with sodium carbonate, were used to raise an antiserum in rabbits. Immunoblot analysis demonstrated the reaction of this antiserum with six peroxisomal integral membrane polypeptides (molecular masses, 140, 69, 50, 36, 22, and 15 kDa). Treatment of rats with the hypolipidemic drug clofibrate caused a 4- to 10-fold induction in the 69-kDa integral membrane polypeptide, while the other integral membrane polypeptides remained unchanged or varied to a lesser extent. The anti-peroxisomal membrane serum reacted with two integral membrane polypeptides of the endoplasmic reticulum which co-migrated with the 50- and 36-kDa integral membrane polypeptides of the peroxisome. Biochemical and immunoblot analyses indicated that these integral membrane polypeptides were co-localized to peroxisomes and endoplasmic reticulum. Immunoprecipitation of in vitro translation products of RNA isolated from free and membrane-bound polysomes indicated that the 22-, 36-, and 69-kDa integral membrane polypeptides were synthesized on free polysomes, while the 50-kDa integral membrane polypeptide was predominantly synthesized on membrane-bound polysomes. The predominant synthesis of the 50-kDa integral membrane polypeptide on membrane-bound polysomes raises interesting possibilities concerning its biosynthesis.
Bodnar
A G
AG
Department of Biochemistry, McMaster University, Hamilton, Ont., Canada.
Rachubinski
R A
RA
D000818:Animals; D015415:Biomarkers; D002458:Cell Fractionation; D002994:Clofibrate / Q000494:pharmacology*; D008099:Liver / Q000737:chemistry / Q000187:drug effects; D008247:Lysosomes / Q000737:chemistry / Q000276:immunology; D008565:Membrane Proteins / Q000737:chemistry* / Q000276:immunology; D008830:Microbodies / Q000737:chemistry* / Q000187:drug effects / Q000276:immunology; D008930:Mitochondria, Liver / Q000737:chemistry / Q000276:immunology; D011233:Precipitin Tests; D014176:Protein Biosynthesis; D051381:Rats; D011919:Rats, Inbred Strains
D015415:Biomarkers; D008565:Membrane Proteins; D002994:Clofibrate