Formation and Nanoscale Characterization of Asymmetric Supported Lipid Bilayers Containing Raft-Like Domains.

The development of new strategies for achieving stable asymmetric membrane models has turned interleaflet lipid asymmetry into a topic of major interest. Cyclodextrin-mediated lipid exchange constitutes a simple and versatile method for preparing asymmetric membrane models without the need for sophisticated equipment. Here we describe a protocol for preparing asymmetric supported lipid bilayers mimicking membrane rafts by cyclodextrin-mediated lipid exchange and the main guidelines for obtaining structural information and quantitative measures of their mechanical properties using Atomic force microscopy and Force spectroscopy; two powerful techniques that allow membrane characterization at the nanoscale.

TBC1D10C is a cytoskeletal functional linker that modulates cell spreading and phagocytosis in macrophages.

Cell spreading and phagocytosis are notably regulated by small GTPases and GAP proteins. TBC1D10C is a dual inhibitory protein with GAP activity. In immune cells, TBC1D10C is one of the elements regulating lymphocyte activation.

Asymmetric bilayers mimicking membrane rafts prepared by lipid exchange: Nanoscale characterization using AFM-Force spectroscopy.

Sphingolipids-enriched rafts domains are proposed to occur in plasma membranes and to mediate important cellular functions. Notwithstanding, the asymmetric transbilayer distribution of phospholipids that exists in the membrane confers the two leaflets different potentials to form lateral domains as next to no sphingolipids are present in the inner leaflet. How the physical properties of one leaflet can influence the properties of the other and its importance on signal transduction across the membrane are questions still unresolved.

Simultaneous determination of the elastic properties of the lipid bilayer by atomic force microscopy: bending, tension, and adhesion.

A nanodrum of an unsupported L-α-phosphatidylcholine bilayer on a ∼7 μm pore was studied using a new experimental setup that permits atomic force microscopy (AFM) in conjunction with the electrical determination of trans-bilayer channels, thus checking its unilamellar character. In these nanodrums, the bilayer engulfs the intruding AFM tip with an adhesion similar to the attraction between two mica supported bilayers brought into close contact. Using this response and the finding of a nonlinear behavior of the Canham-Helfrich elastic model allows for the simultaneous determination of the elastic properties of the membrane.