Effect of actin concentration on the structure of actin-containing liposomes

Liposomes encapsulating monomeric actin (G-actin) were produced via extrusion through 400 and 600 nm pore diameter polycarbonate membranes in low ionic strength buffer (G-buffer). After actin-containing liposomes were subjected to high ionic strength polymerization buffer (F-buffer), morphological changes in the structure of actin-containing liposomes were studied using asymmetric flow field-flow fractionation (AFFF) coupled with multiangle static light scattering (MASLS). The general shape of these liposomes was initially determined by fitting three form factors, which describe the angular distribution of scattered light from a spherical thin shell, thin disk, and thin rod, to the experimentally measured light scattering spectra to regress the dimensions of the liposomes corresponding to the proposed geometry. Light scattering spectra that yielded a best fit to the thin disk model were analyzed further and fit to the ellipsoidal of revolution form factor to regress both the major and minor axis dimensions. The results of this analysis showed that actin-containing liposomes extruded through 400 and 600 nm pore diameter membranes in F-buffer, at a low actin concentration (0.1 mg/mL), assumed a spherical shape, which is also the case for plain liposomes (no actin present) in G-buffer and F-buffer. When the actin concentration was increased to 1 mg/mL, the polymerizing actin filaments stretched the initially spherical liposome into a disklike shape. However, when the actin concentration was further increased to 5 mg/mL, the liposomes reverted back to a spherical shape.