1 research outputs found
A Programmable DNA Origami Platform to Organize SNAREs for Membrane Fusion
Soluble <i>N</i>-ethylmaleimide-sensitive factor attachment protein receptor (SNARE)
complexes are the core molecular machinery of membrane fusion, a fundamental
process that drives inter- and intracellular communication and trafficking.
One of the questions that remains controversial has been whether and
how SNAREs cooperate. Here we show the use of self-assembled DNA-nanostructure
rings to template uniform-sized small unilamellar vesicles containing
predetermined maximal number of externally facing SNAREs to study
the membrane-fusion process. We also incorporated lipid-conjugated
complementary ssDNA as tethers into vesicle and target membranes,
which enabled bypass of the rate-limiting docking step of fusion reactions
and allowed direct observation of individual membrane-fusion events
at SNARE densities as low as one pair per vesicle. With this platform,
we confirmed at the single event level that, after docking of the
templated-SUVs to supported lipid bilayers (SBL), one to two pairs
of SNAREs are sufficient to drive fast lipid mixing. Modularity and
programmability of this platform makes it readily amenable to studying
more complicated systems where auxiliary proteins are involved