Aggregation and fusion of phospholipid vesicles

Abstract

Membrane fusion and aggregation of phospholipid vesicles are reviewed and discussed. The fusion process is viewed as consisting of several stages: aggregation and close apposition of the particles, destabilization, and finally, merging of the bilayers. A procedure is presented which yields both the rate constant of the dimerization (C 11) and the rate constant for fusion of the dimers (f 11), which is a direct measure of the probability that two apposed vesicles will fuse. Experimental methods used in the study of membrane fusion are reviewed, primarily with respect to their capacity to monitor the kinetics of vesicle fusion. A few kinetic studies on the mixing of aqueous contents, leakage and increase in size of fusing vesicles are presented in detail. The range of C 11 values for Ca 2+-induced aggregation and fusion of small unilamellar vesicles (SUV, ∼ 125 Å radius) composed of phosphatidylserine (PS) is 10 6 to 5 × 10 7 M -1 in the presence of Ca 2+ concentrations from 1.15 to 2 mM, respectively. For larger PS vesicles (LUV, ∼ 500 Å radius) C 11 = 6.5 × 10 7 M -1s -1 in the presence of 5 mM Ca 2+. These values are in good agreement with theoretical calculations based on van der Waals and electrostatic interactions, in which binding of cations is explicitly taken into account. The rate constants of fusion, f 11, are 5 s -1 for PS SUV and 0.08 s -1 for LUV in the presence of 2 mM and 5 mM Ca 2+, respectively. The significance of these fusion rate constants to the duration of the fusion event is discussed. Factors affecting fusion such as cations, temperature, membrane composition vesicle concentration and size are reviewed and analyzed. Di- or tri-valent cations induce fusion of acidic phospholipid vesicles (except for phosphatidylinositol) in either pure or mixed form. Among the neutral phospholipids, phosphatidylcholine (PC) inhibits but phosphatidylethanolamine (PE) sustains or enhances the fusion capacity of acidic phospholipid vesicles. Monovalent cations induce reversible aggregation of negatively charged vesicles, but they inhibit the fusion induced by divalent cations such as Ca 2+ or Mg 2+. Fusion of neutral phospholipid vesicles, and it occurs the cation-induced fusion of acidic phospholipid vesicles, and it occurs only at temperatures below the gel to liquid crystalline phase transition temperature Tc. This is in contrast to the acidic phospholipid vesicle fusion which is greatly enhanced when the temperature is above the Tc of the phospholipid.