Atomic force microscopy and light scattering of small unilamellar actin-containing liposomes.
Three-dimensional networks of filamentous actin (F-actin) encapsulated inside phosphatidylcholine liposomes are currently being used in an effort to model the cytoskeleton and plasma membrane of eukaryotic cells. In this article, unilamellar lipid vesicles consisting of egg yolk-derived phosphatidylcholine encapsulating monomeric actin (G-actin) were made via extrusion in low ionic strength buffer (G-buffer). Vesicle shape and structure in these dispersions was studied using a combination of fluid-tapping atomic force microscopy, and multiangle static light scattering. After subjecting the liposome dispersion to high ionic strength polymerization buffer (F-buffer) containing K(+) ions, atomic force microscopy imaging and light scattering of these liposomes indicated the formation of specialized structures, including an overall liposome structure transformation from spherical to torus, disk-shaped geometries and tubular assemblies. Several atomic force microscopy control measurements were made to ascertain that the specialized structures formed were not due to free G-actin and F-actin self-assembling on the sample surface, plain liposomes exposed to G- and F-buffer, or liposomes encapsulating G-actin. Liposomes encapsulating G-actin assumed mostly thin disk shapes and some large irregularly shaped aggregates. In contrast, liposomes encapsulating polymerized actin assumed mostly torus or disk shapes along with some high aspect ratio tubular structures.
Palmer
Andre F
AF
Department of Chemical and Biomolecular Engineering, Center for Molecularly Engineered Materials, University of Notre Dame, Notre Dame, Indiana 46556, USA. apalmer@nd.edu
Wingert
Philip
P
Nickels
Jonathan
J
D000199:Actins / Q000737:chemistry* / Q000648:ultrastructure*; D040761:Biomimetic Materials / Q000737:chemistry*; D008027:Light; D008081:Liposomes / Q000737:chemistry*; D046911:Macromolecular Substances; D008560:Membrane Fluidity; D008567:Membranes, Artificial; D018625:Microscopy, Atomic Force / Q000379:methods*; D012031:Refractometry / Q000379:methods*; D012542:Scattering, Radiation; D013499:Surface Properties
D000199:Actins; D008081:Liposomes; D046911:Macromolecular Substances; D008567:Membranes, Artificial