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Free keywords:
GUV; intraluminal vesicles; bending rigidity; membrane tension; protein domain; microfluidics; micropipette aspiration
Abstract:
The endosomal sorting complex required for transport (ESCRT) is a multi-protein machinery involved in several membrane remodelling processes. Different approaches have been used to resolve how ESCRT proteins scission membranes. However, the underlying mechanisms generating membrane deformations are still a matter of debate. Here, giant unilamellar vesicles, microfluidic technology and micropipette aspiration are combined to continuously follow the ESCRT-III-mediated membrane remodelling on the single-vesicle level for the first time. With this approach, we identify different mechanisms by which a minimal set of three ESCRT-III proteins from Entamoeba histolytica reshape the membrane. These proteins modulate the membrane stiffness and spontaneous curvature to regulate bud size and generate intraluminal vesicles even in the absence of ATP. We show that the bud stability depends on the protein concentration and membrane tension. The approaches introduced here should open the road to diverse applications in synthetic biology for establishing artificial cells with several membrane compartments.
