Real-Time QCM-D Monitoring of Electrostatically Driven Lipid Transfer between Two Lipid Bilayer Membranes

Abstract

The lipid exchange/transfer between lipid membranes is important for many biological functions. To learn more about how the dynamics of such processes can be studied, we have investigated the interaction of positively and negatively charged lipid vesicles with supported lipid bilayers (SLBs) of opposite charge. The vesicle-SLB interaction leads initially to adsorption of lipid vesicles on the SLB, as deduced from the mass uptake kinetics and the concerted increase in dissipation, monitored by the quartz crystal microbalance with dissipation (QCM-D) technique. Eventually, however, vesicles (and possibly other lipid structures) desorb from the SLB surface, as judged from the mass loss and the dissipation decrease. The mass loss is approximately as large as the initial mass increase; i.e., at the end of the process the mass load is that of a SLB. We interpret this interesting kinetics in terms of initial strong electrostatic attraction between the added vesicles and the SLB, forming a structure where lipid transfer between the two bilayers occurs on a time scale of 10-40 min. We suggest that this lipid transfer causes a charge equilibration with an accompanying weakening of the attraction, and eventually repulsion, between the SLB and vesicles, leading to desorption of vesicles from the SLB. The composition of the latter has thus been modified compared to the initial one, although no net mass increase or decrease has occurred. Direct evidence for the lipid exchange was obtained by sequential experiments with alternating positive and negative vesicles, as well as by using fluorescently labeled lipids and FRAP. The above interpretation was further strengthened by combined QCM-D and optical reflectometry measurements.