Multiplicity of antioxidant enzyme catalase in mouse liver cells.
Multiplicity of catalase activity has been observed in crude homogenates from the tissue and cell lines of mouse liver by ethanol/Triton X-100/heat treatment. The five enzymatically active catalase bands were designated as CAT1, CAT2, CAT3, CAT4, and CAT5 with a nondenatured molecular mass of 270kDa, 258kDa, 229kDa, 210kDa, or 197kDa, respectively. Cultured mouse liver cell lines, mouse liver tissue homogenate, and pure mouse liver catalase showed only one catalase band (CAT1) after ethanol/Triton X-100 treatment at 4 degrees C for 72 hr. The same treatment but incubated at 37 degrees C for 72 hr yielded three bands (CAT2, CAT4, CAT5) in normal cell line, only one band (CAT5) in MNNG-transformed and SV40-transformed cells, two bands (CAT1, CAT4) in mouse liver tissue homogenates, and two bands (CAT1, CAT3) in pure mouse liver catalase. These five catalase bands were further biochemically characterized. The CAT1, CAT2, and CAT3 are sensitive to heat (68 degrees C, 1 min), while CAT4 and CAT5 are rather heat resistant. The sensitivity to catalase inhibitors, such as aminotriazole, azide, or cyanide varies among the isoforms. Protease inhibitors could prevent the formation of CAT3 and CAT4, but not CAT5. Treatment with protease, however, removed all forms of catalase except CAT5. We conclude from this study that the appearance of different catalase bands is likely due to epigenetic modification of the protein, particularly proteolysis. The lowered catalase activity in transformed cells might also be attributable to the loss of two catalase isoforms.
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Radiation Research Laboratory, University of Iowa, Iowa City 52242, USA.
D000818:Animals; D000975:Antioxidants / Q000378:metabolism*; D002374:Catalase / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D002461:Cell Line, Transformed; D010450:Endopeptidases / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004795:Enzyme Stability; D004912:Erythrocytes / Q000201:enzymology; D000431:Ethanol / Q000494:pharmacology; D005979:Glutathione Peroxidase / Q000097:blood; D006358:Hot Temperature; D006801:Humans; D007527:Isoenzymes / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D008099:Liver / Q000201:enzymology*; D008769:Methylnitronitrosoguanidine; D051379:Mice; D008970:Molecular Weight; D017830:Octoxynol / Q000494:pharmacology; D011480:Protease Inhibitors / Q000494:pharmacology; D013539:Simian virus 40
D000975:Antioxidants; D004791:Enzyme Inhibitors; D007527:Isoenzymes; D011480:Protease Inhibitors; D008769:Methylnitronitrosoguanidine; D000431:Ethanol; D017830:Octoxynol; D002374:Catalase; D005979:Glutathione Peroxidase; D010450:Endopeptidases