Interactions between the proteins of the ternary soluble N-ethyl maleimide-sensitive fusion protein attachment protein receptor (SNARE) complex, synaptobrevin 2 (Sb2), syntaxin 1A (Sx1A) and synaptosome-associated protein of 25kDa (SNAP25) can be readily assessed using force spectroscopy single-molecule measurements. We studied interactions during the disassembly of the ternary SNARE complex pre-formed by binding Sb2 in parallel or anti-parallel orientations to the binary Sx1A-SNAP25B acceptor complex. We determined the spontaneous dissociation lifetimes and found that the stability of the anti-parallel ternary SNARE complex is ~1/3 less than that of the parallel complex. While the free energies were very similar, within 0.5 kBT, for both orientations, the enthalpy changes (42.1 kBT and 39.8 kBT, for parallel and anti-parallel orientations, respectively) indicate that the parallel ternary complex is energetically advantageous by 2.3 kBT. Indeed, both ternary SNARE complex orientations were much more stable (by ~4-13 times) and energetically favorable (by ~9-13 kBT) than selected binary complexes, constituents of the ternary complex, in both orientations. We propose a model which considers the geometry for the vesicle approach to the plasma membrane with favorable energies and stability as the basis for preferential usage of the parallel ternary SNARE complex in exocytosis. © 2012 Elsevier Ltd.