Partition of catalase and its peroxidase activities in human red cell membrane. Effect of ATP depletion

L. M. Snyder, S. C. Liu, J. Palek, P. Bulat, L. Edelstein, S. K. Srivastava, N. L. Fortier

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Abstract

Partition of catalase (hydrogen-peroxide:hydrogen-peroxide oxidoreductase EC 1.11.1.6) and peroxidase (donor:hydrogen-peroxide oxidoreductase EC 1.11.1.7) activities between the red cell membrane and the cytosol were studied under various experimental conditions. A small but significant amount of catalase (1.6%) was retained on human red cell membranes prepared by hemolysing washed red cells with 30 volumes of 10 mM Tris buffer, pH 7.4. Membrane-bound catalase had a relatively higher peroxidase activity than the soluble enzyme fraction. Polyacrylamide gel electrophoresis in sodium dodecyl sulfate of the solubilized membranes demonstrated catalase to be a single band with a molecular weight of 60 000. Membranes prepared from adenosine triphosphate-depleted red cells depicted a two to three-fold increase in catalase activity, as well as an increase in 60 000 molecular weight band on polyacrylamide gel electrophoresis. The extra amount of retained catalase was a less efficient peroxidase than found in fresh membranes. The binding of catalase to ATP-depleted red cell membranes was dependent upon both pH and hemolysing ratio. Red cells incubated at pH 7.1 demonstrated a decrease in bound catalase, as did membranes prepared from red cells hemolysed at 1:100 dilution. β-Mercaptoethanol decreased the catalase activity in the membranes and increased the o-dianisidine peroxidase activity without any significant effect on the 60 000-dalton band.

Original languageEnglish (US)
Pages (from-to)290-302
Number of pages13
JournalBBA - Biomembranes
Volume470
Issue number2
DOIs
StatePublished - Oct 17 1977

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ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Cell Biology

Cite this

Snyder, L. M., Liu, S. C., Palek, J., Bulat, P., Edelstein, L., Srivastava, S. K., & Fortier, N. L. (1977). Partition of catalase and its peroxidase activities in human red cell membrane. Effect of ATP depletion. BBA - Biomembranes, 470(2), 290-302. https://doi.org/10.1016/0005-2736(77)90107-9