Abstract
A single lipid vesicle can be regarded as an autonomous ultra-miniaturised 3D biomimetic “scaffold” (Ø ≥ 13 nm) ideally suited for reconstitution and interrogation of biochemical processes. The enclosing lipid bilayer membrane of a vesicle can be applied for studying binding (protein/lipid or receptor/ligand interactions) or transmembrane events (membrane permeability or ion channel activation) while the aqueous vesicle lumen can be used for confining few or single macromolecules and probe, e.g., protein folding, catalytic pathways of enzymes or more complex biochemical reactions, such as signal transduction cascades. Immobilisation (arraying) of single vesicles on a solid support is an extremely useful technique that allows detailed characterisation of vesicle preparations using surface sensitive techniques, in particular fluorescence microscopy. Surface-based single vesicle arrays allow a plethora of prototypic sensing applications in a high throughput format with high spatial and high temporal resolution. In this review we present a series of applications of single vesicle arrays for screening/sensing of: membrane curvature dependent protein-lipid interactions, bilayer tension, reactions triggered in the vesicle lumen, the activity of transmembrane protein channels and biological membrane fusion reactions.
Highlights
A lipid vesicle is comprised by an aqueous lumen confined by a lipid bilayer envelope
The review is structured as a series of examples on screening/sensing of: curvature dependent protein-lipid interactions, bilayer tension, reactions triggered in the vesicle lumen, the activity of transmembrane protein channels and biological membrane fusion reactions
Bränden et al [40] in an elegant piece of work demonstrated quantitative measurements of the permeability properties of an aquaglyceroporin using evanescent-wave sensing. This was accomplished by immobilising a layer of vesicles carrying the reconstituted porin on a SPR compatible surface, which permitted label-free probing of the efflux kinetics of sugar alcohols into the immobilised vesicles
Summary
A lipid vesicle is comprised by an aqueous lumen confined by a lipid bilayer envelope. A vesicle can be readily derivatised with organic and inorganic moieties creating an autonomous complex system of nanoscopic dimensions. This biomimetic “scaffold” can host motifs that permit surface immobilisation of the vesicle, identity encoding, monitoring of biorecognition processes taking place at or within the bilayer, probing of transmembrane transport and recording of biochemical reactions in general, see Figure 1(a). We focus on applications of surface-based arrays of single lipid vesicles. The review is structured as a series of examples on screening/sensing of: curvature dependent protein-lipid interactions, bilayer tension, reactions triggered in the vesicle lumen, the activity of transmembrane protein channels and biological membrane fusion reactions
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