Kinetic analysis of fusion of hemagglutinating virus of Japan with erythrocyte membrane using spin-labeled phosphatidylcholine

Abstract

HVJ* (hemagglutinating virus of Japan containing spin-labeled phosphatidylcholine in its envelope around 10 mol %) was adsorbed onto erythrocytes or erythrocyte ghosts at various doses, and the ESR spectrum of the virus-cell system was measured at 37.degree. C. The peak-height increase for the HVJ*-ghost system was satisfactorily analyzed on the basis of envelope fusion by a first-order kinetic equation with two different rate constants. The rate constant was obtained as k1 = 0.84 min-1 and k2 = 0.011 min-1, independent of the virus dose. The fraction of virus fused at the rate constant k1 decreased with the dose. However, the average number of fast-fusing viruses per cell was nearly independent of the dose, and the value was one to two. The peak-height increase in the HVJ*-erythrocyte system was caused by both envelope fusion and phospholipid exchange catalyzed by the virus-induced hemolyzate. At lower doses, where the virus-induced hemolysis was small and, therefore, the rate of phospholipid exchange was small, the peak-height increase could be analyzed by the same kinetic equation with nearly the same rate constant value for k1 as that for HVJ*-ghosts. However, the k2 was larger than that for HVJ*-ghost, owing to the additional transfer by phospholipid exchange.