EFFECTS OF SULFHYDRYL INHIBITION ON RED BLOOD CELLS. I. MECHANISM OF HEMOLYSIS*

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Abstract
The addition to human red cells of the freely permeable sulfhydryl inhibitor, N-ethylmaleimide, caused the following sequence of events: 1) intracellular glutathione was blocked; 2) glycolysis was inhibited; 3) cation gradients across the cell membrane were disrupted, with a loss of cellular K+ and a gain of Na+ and water; and 4) the influx of water caused by the unopposed oncotic activity of hemoglobin and other macromolecules led to spherocytosis and swelling of the cells, which resulted eventually in the leakage of hemoglobin (i.e., hemolysis). Comparative studies of immune hemolysis revealed that the lysis of red cells by complement does not occur by a comparable osmotic mechanism. The addition to red cells of the nonpermeably sulfhydryl inhibitor, p-mercuribenzoate, also disrupted cation gradients and thereby caused spherocytosis and osmotic hemolysis; however, this inhibitor produced such alterations without blocking intracellular glutathione or inhibiting over-all glycolysis. It is concluded that the shape and viability of red cells depend primarily upon membrane, and not upon intracellular, sulfhydryl activity.