Quantitative characterization of neuronal trans-SNARE complexes using DNA origami

Project Summary/Abstract A key step in neurotransmission is the fusion of the synaptic vesicle (SV) membrane with neuronal plasma membrane (PM), to release neurotransmitters into the synaptic cleft where they bind and activate post synaptic receptors.

A protein complex called SNARE is believed to play a central role since its assembly can generate enough energy to drive fusion.

The current hypothesis that describes SNARE-mediated fusion is referred to as 'SNARE zippering': a v-SNARE protein on SV binds to a t-SNARE protein heterodimer on PM in a zipper-like fashion, forming a trans-SNARE complex (i.e. v- and t-SNARE transmembrane domains are embedded in separate membranes); the released energy eventually overcomes the repulsive forces between SV and PM and pulls the two membranes together, where trans-SNARE complexes transform into cis-SNARE complexes (i.e. v- and t-SNAREs locate on a single membrane).

At present most of what is known concerning neuronal SNARE structure and dynamics stems from analysis of cis-SNARE, but the 'real hero' trans-SNARE that provides the driving force for membrane fusion remains elusive.

A main technical challenge here is to capture partially assembled trans-SNARE complexes that form during the fast process of exocytosis (