Catalase in guinea pig hepatocytes is localized in cytoplasm, nuclear matrix and peroxisomes.
We have compared the intracellular localization of catalase and another peroxisomal marker enzyme, alpha-hydroxy acid oxidase (HAOX), in the livers of guinea pig and rat using immunoelectron microscopy and subcellular fractionation combined with immunoblotting and enzyme activity determination. Antibodies against both enzymes were raised in rabbits and their specificities established by immunoblotting. By immunoelectron microscopy, gold particles representing antigenic sites for catalase were found in guinea pig hepatocytes not only in peroxisomes but also in the cytoplasm and the nuclear matrix. In rat liver, however, catalase was localized exclusively in peroxisomes with no cytoplasmic labeling. Moreover, in both species HAOX was found only in peroxisomes. Subcellular fractionation revealed that purified peroxisomes from both species contained comparable levels of each, catalase and HAOX activities. The total catalase activity, however, was substantially higher in guinea pig and most of this excess catalase was in the cytosolic fraction with some activity also in nuclei. In rat liver, 30 to 40% of both enzymes and in guinea pig liver 30% of HAOX were recovered in the supernatant fraction implying that the fragility of peroxisomes in both species is quite comparable. These observations establish the occurrence of extraperoxisomal catalase in guinea pig liver. The catalase in the cytoplasm and nucleus of liver parenchymal cells is most probably involved in scavenging of H2O2, protecting the cell against toxic and mutagenic effects of this noxious agent.
Yamamoto
K
K
Department of Anatomy, University of Heidelberg, Federal Republic of Germany.
Völkl
A
A
Hashimoto
T
T
Fahimi
H D
HD
D000818:Animals; D000936:Antigen-Antibody Complex / Q000032:analysis; D002374:Catalase / Q000378:metabolism*; D002458:Cell Fractionation; D002467:Cell Nucleus / Q000201:enzymology* / Q000648:ultrastructure; D003593:Cytoplasm / Q000201:enzymology / Q000648:ultrastructure; D005260:Female; D006168:Guinea Pigs; D008099:Liver / Q000201:enzymology* / Q000648:ultrastructure; D008830:Microbodies / Q000201:enzymology* / Q000648:ultrastructure; D008854:Microscopy, Electron; D051381:Rats; D011919:Rats, Inbred Strains
D000936:Antigen-Antibody Complex; D002374:Catalase