Abstract
Amphiphilic peptides such as mastoparan (MP) lyses erythrocytes, but the mechanism of lytic effect of MP are still unclear. In the present study, we have developed a novel flow injection analysis system to elucidate the dynamics of hemolysis. This system is composed of a flow pump for HPLC, autoinjector, cell-retaining chamber, detector and data processor. At first, erythrocytes from male Wistar rats were injected into a cellcetaining chamber where the erythrocytes were kept on a membrane filter. Hemolytic stimulation was given to the erythrocytes through the autoinjector, and hemoglobin released into the flow stream from erythrocytes was monitored photometrically at 413 nm. This system had a high precision and correlation (r = 0.989) with the bath application method. Using this system, the hemolytic action of MP was analyzed. The hemolytic activity of MP depended upon the number of MP molecules, not upon the reaction time. MP-induced hemolysis proceeded promptly and transiently. The mechanism of MP-induced hemolysis might be different from that of the SDS-induced hemolysis. Because our flow injection analysis system can be anto- mated easily, it will be useful for the clinical or basic research
