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J. Biochem, 1974, Vol. 76, No. 4 703-708
© 1974 Japanese Biochemical Society

research-article

Studies on Rat Liver Catalase

VII. Double-labeling of Catalase by 14C-Leucine and 3H-{delta}-Aminolevulinic Acid*

Tokuhiko HIGASHI, Fumiko KAWAMATA and Terufumi SAKAMOTO

Department of Biochemistry, School of Pharmaceutical Sciences, Showa University Shinagawa-ku, Tokyo

To investigate the mechanism of incorporation of heme into newly-synthesized catalase [EC 1.11.1.6 [EC] ] rat liver catalase was labeled simultaneously with 14C-leucine and 3H-{delta}-aminolevulinic acid both in vivo and in vitro. The results obtained were as follows:

  1. The kinetics of incorporation of 3H of injected {delta}-aminolevulinic acid into catalase on ribosomes were the same as those of incorporation of 14C-leucine.
  2. In a cell-free system newly-synthesized catalase was found to be labeled with both isotopes.
  3. The ratio of 3H/14C incorporated in vivo was lowest in catalase of microsomes and ribosomes, highest in that of peroxisomes, and intermediate in catalase of the soluble fraction of liver cells. The former remained almost constant, while the latter two increased to a plateau after injection of the isotopic tracers.

Results suggested that an intermediate is formed which contains less heme than in the completed molecule. Further addition of heme to this intermediate seems to occur in the cytosol, and probably also in the peroxisomes.

*This work was presented at the 46th Annual Meeting of the Japanese Biochemical Society (Nagoya, September 27, 1973).


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