Journal of Structural Biology, Vol.184, No.1, 43-51, 2013
Clathrin-coated vesicles from brain have small payloads: A cryo-electron tomographic study
Clathrin coats, which stabilize membrane curvature during endocytosis and vesicular trafficking, form highly polymorphic fullerene lattices. We used cryo-electron tomography to visualize coated particles in isolates from bovine brain. The particles range from similar to 66 to similar to 134 nm in diameter, and only 20% of them (all >= 80 nm) contain vesicles. The remaining 80% are clathrin "baskets", presumably artifactual assembly products. Polyhedral models were built for 54 distinct coat geometries. In true coated vesicles (CVs), most vesicles are offset to one side, leaving a crescent of interstitial space between the coat and the membrane for adaptor proteins and other components. The latter densities are fewer on the membrane-proximal side, which may represent the last part of the vesicle to bud off. A small number of densities - presumably cargo proteins - are associated with the interior surface of the vesicles. The clathrin coat, adaptor proteins, and vesicle membrane contribute almost all of the mass of a CV, with most cargoes accounting for only a few percent. The assembly of a CV therefore represents a massive biosynthetic effort to internalize a relatively diminutive payload. Such a high investment may be needed to overcome the resistance of membranes to high curvature. Published by Elsevier Inc.
Keywords:Clathrin-mediated endocytosis;Cryo-electron microscopy;Three-dimensional image reconstruction;Adaptor proteins;Fullerenes