Investigation of peroxisomal lipid beta-oxidation enzymes in guinea pig liver peroxisomes by immunoblotting and immunocytochemistry.
We investigated the immunoreactivity of the peroxisomal lipid beta-oxidation enzymes acyl-CoA oxidase, trifunctional protein, and thiolase in guinea pig liver and compared it with that of homologous proteins in rat, using immunoblotting of highly purified peroxisomal fractions and monospecific antibodies to rat proteins. In addition, the immunocytochemical localization of beta-oxidation enzymes in guinea pig liver was compared with that of catalase. All antibodies showed crossreactivity between the two species, indicating that these peroxisomal proteins have been well conserved, although all exhibited some differences with respect to molecular size and, in the case of acyl-CoA oxidase, in frequency of the immunoreactive bands. In the latter case, a distinct second band in the 70 KD range was observed in guinea pig, in addition to the regular band due to subunit A present in rat liver. This novel band could be due either to trihydroxycoprostanoyl-CoA oxidase or to the non-inducible branched chain fatty acid oxidase described recently. All three beta-oxidation enzymes were immunolocalized by light and electron microscopy to the matrix of peroxisomes, in contrast to catalase, which is also found in the cytoplasm and the nucleus of hepatocytes in guinea pig liver.
Department of Anatomy and Cell Biology II, University of Heidelberg, Germany.
K
K
Yamamoto
A
A
Völkl
H D
HD
Fahimi
D015094:3-Hydroxyacyl CoA Dehydrogenases / Q000032:analysis*; D000101:Acetyl-CoA C-Acetyltransferase / Q000032:analysis*; D042962:Acyl-CoA Oxidase; D000818:Animals; D000918:Antibody Specificity; D002374:Catalase / Q000032:analysis; D002467:Cell Nucleus / Q000201:enzymology; D003593:Cytoplasm / Q000201:enzymology; D004746:Enoyl-CoA Hydratase / Q000032:analysis*; D005260:Female; D006168:Guinea Pigs; D015151:Immunoblotting; D007150:Immunohistochemistry; D007535:Isomerases / Q000032:analysis*; D008099:Liver / Q000166:cytology / Q000201:enzymology / Q000648:ultrastructure*; D008297:Male; D008830:Microbodies / Q000201:enzymology* / Q000648:ultrastructure; D016253:Microscopy, Immunoelectron; D008970:Molecular Weight; D009097:Multienzyme Complexes / Q000032:analysis*; D010088:Oxidoreductases / Q000032:analysis*; D063994:Peroxisomal Bifunctional Enzyme; D051381:Rats; D013045:Species Specificity
D009097:Multienzyme Complexes; D010088:Oxidoreductases; D015094:3-Hydroxyacyl CoA Dehydrogenases; D002374:Catalase; D042962:Acyl-CoA Oxidase; D000101:Acetyl-CoA C-Acetyltransferase; D004746:Enoyl-CoA Hydratase; D063994:Peroxisomal Bifunctional Enzyme; D007535:Isomerases