title: Continuous lipid bilayers derived from cell membranes for spatial molecular manipulation.
abstract: Progress with respect to enrichment and separation of native membrane components in complex lipid environments, such as native cell membranes, has so far been very limited. The reason for the slow progress can be related to the lack of efficient means to generate continuous and laterally fluid supported lipid bilayers (SLBs) made from real cell membranes. We show in this work how the edge of a hydrodynamically driven SLB can be used to induce rupture of adsorbed lipid vesicles of compositions that typically prevent spontaneous SLB formation, such as vesicles made of complex lipid compositions, containing high cholesterol content or being derived from real cell membranes. In particular, upon fusion between the moving edge of a preformed SLB and adsorbed vesicles made directly from 3T3 fibroblast cell membranes, the membrane content of the vesicles was shown to be efficiently transferred to the SLB. The molecular transfer was verified using cholera toxin B subunit (CTB) binding to monosialoganglioside receptors (G(M1) and G(M3)), and the preserved lateral mobility was confirmed by spatial manipulation of the G(M1/M3)-CTB complex using a hydrodynamic flow. Two populations of CTB with markedly different drift velocity could be identified, which from dissociation kinetics data were attributed to CTB bound with different numbers of ganglioside anchors.
authors.affiliation: Department of Applied Physics, Chalmers University of Technology, Gothenburg, Sweden.
authors.forename: Lisa
authors.initials: L
authors.lastname: Simonsson
authors.forename: Anders
authors.initials: A
authors.lastname: Gunnarsson
authors.forename: Patric
authors.initials: P
authors.lastname: Wallin
authors.forename: Peter
authors.initials: P
authors.lastname: Jönsson
authors.forename: Fredrik
authors.initials: F
authors.lastname: Höök
mesh_terms: D016475:3T3 Cells; D000818:Animals; D002462:Cell Membrane; D002772:Cholera Toxin / Q000737:chemistry / Q000378:metabolism*; D002784:Cholesterol / Q000378:metabolism; D005347:Fibroblasts / Q000737:chemistry; D057446:Hydrodynamics; D008051:Lipid Bilayers / Q000737:chemistry / Q000378:metabolism*; D008561:Membrane Fusion; D051379:Mice; D008846:Micromanipulation / Q000379:methods*; D011485:Protein Binding; D011956:Receptors, Cell Surface / Q000737:chemistry / Q000378:metabolism*
chemical_list: D008051:Lipid Bilayers; D011956:Receptors, Cell Surface; C034993:ganglioside receptor; D002772:Cholera Toxin; D002784:Cholesterol