ERYTHROCYTE METABOLISM. VI. SEPARATION OF ERYTHROCYTE ENZYMES FROM HEMOGLOBIN*

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Abstract
A method, using DEAE-cellulose, is described for the isolation of a hemoglobin-free, enzyme protein fraction from erythrocyte hemolysates. The isolated fraction contains approximately 140 mg of protein derived from 10 ml of packed red cells. In addition, the presence of ATP and ADP and small amounts of other nucleotides is demonstrated. The absorbancy at 260 m[mu] in the enzyme fraction can be accounted for by the nucleotides and that at 410 m[mu] is attributed to the heme components of catalase and TPNH-methemoglobin reductase. Various glycolytic enzymes and a number of miscellaneous enzymes (catalase, TPNH-methemoglobin reductase, nucleoside phosphorylase, glucose-6-phosphate dehydrogenase, glutathione reductase, adenylic deaminase and formate-activating enzyme) are recovered almost quantitatively in the enzyme protein fraction.