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 language | English (US) |
|---|---|
| Pages (from-to) | 290-302 |
| Number of pages | 13 |
| Journal | BBA - Biomembranes |
| Volume | 470 |
| Issue number | 2 |
| DOIs | |
| State | Published - Oct 17 1977 |
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ASJC Scopus subject areas
- Biochemistry
- Biophysics
- Cell Biology
- Medicine(all)
Cite this
Partition of catalase and its peroxidase activities in human red cell membrane. Effect of ATP depletion. / Snyder, L. M.; Liu, S. C.; Palek, J.; Bulat, P.; Edelstein, L.; Srivastava, Satish; Fortier, N. L.
In: BBA - Biomembranes, Vol. 470, No. 2, 17.10.1977, p. 290-302.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Partition of catalase and its peroxidase activities in human red cell membrane. Effect of ATP depletion
AU - Snyder, L. M.
AU - Liu, S. C.
AU - Palek, J.
AU - Bulat, P.
AU - Edelstein, L.
AU - Srivastava, Satish
AU - Fortier, N. L.
PY - 1977/10/17
Y1 - 1977/10/17
N2 - 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.
AB - 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.
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U2 - 10.1016/0005-2736(77)90107-9
DO - 10.1016/0005-2736(77)90107-9
M3 - Article
C2 - 20953
AN - SCOPUS:0017669518
VL - 470
SP - 290
EP - 302
JO - Biochimica et Biophysica Acta - Biomembranes
JF - Biochimica et Biophysica Acta - Biomembranes
SN - 0005-2736
IS - 2
ER -