Synexin enhances the aggregation rate but not the fusion rate of liposomes

The effect of synexin on the calcium-induced fusion of large unilamellar liposomes was studied by using two assays for the mixing of aqueous contents. The results were analyzed in terms of the mass action kinetic model, which describes the overall fusion reaction as a two-step sequence consisting of a second-order process of liposome aggregation followed by a first-order fusion reaction. By using several different lipid compositions and varying the electrolyte composition, it was possible to select the rate-limiting step of the overall fusion process. When aggregation was the rate-limiting step, as in the case of Ca2+-induced fusion of phosphatidylserine (PS), phosphatidate (PA)/phosphatidylethanolamine (PE) (1:3), and PS/PE (1:3) liposomes, synexin increased the overall fusion kinetics by increasing the aggregation rate constant (up to 100-fold). When aggregation was rapid compared to destabilization of apposed membranes, i.e., fusion was rate limiting, synexin either had no effect or reduced the overall fusion kinetics. In one such case involving liposomes composed of PA/PS/PE/phosphatidylcholine (PC) (10:15:65:10), synexin reduced the fusion rate constant by 50%. The effect of calcium-induced synexin polymerization was investigated by preincubation of synexin with calcium prior to addition of liposomes. Prepolymerization by Ca2+ always decreased the activity of synexin such that it was less than the activity of an equal amount of untreated monomers. However, it was found that the activity of synexin monomers polymerized to an average hexameric size was greater than that of one-sixth as many untreated monomers, with respect to the liposome aggregation rate constant. Neither polymers nor monomers increased the fusion rate constant.