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A handful of recent reports concluded that complexins clamp SNARE-catalyzed membrane fusion (4− 8) and that Ca2+-syt displaces complexin from SNARE complexes to alleviate the clamp (4− 6, 8, 13).
For the purpose of proteins bio-conjugation to Gold Nano-Particles (GNPs), we designed and synthesized a polypeptide named NanoLock, derived from SNARE (Soluble NSF Attachment protein REceptor) proteins and able to form a remarkably stable complex with the SNARE protein SNAP25 (Synaptosome Associated Protein of 25 kDa).
It has been proposed that complexin clamps membrane fusion and that Ca2+-synaptotagmin displaces complexin from SNARE complexes to relieve this clamping activity.
Unbound SNAREs were removed from SNARE complexes by washing the resins with SM assay buffer twice at 4°C and twice at room temperature.
However, it has also been reported that complexins are not displaced from SNARE complexes by syt (6, 9) and that complexins are not clamps, but instead play a positive role in SNARE-mediated fusion (10− 10−.
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A mismatch between the force output from SNAREs and the load from membranes would inevitably lead to dissipation of the SNARE zippering energy into heat, reducing the efficiency or the rate of membrane fusion.
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