Snapshot of sequential SNARE assembling states between membranes shows that N-terminal transient assembly initializes fusion

Yong Jian Wang (1) , Feng Li (2) , Nicolas Rodriguez (3) , Xavier Lafosse (4) , Christine Gourier (1) , Eric Perez (1) , Frederic Pincet (1)

1 LPS - Laboratoire de Physique Statistique de l'ENS
2 Department of Cell Biology
3 UPMC - Université Pierre et Marie Curie - Paris 6
4 LPN - Laboratoire de photonique et de nanostructures

Many prominent biological processes are driven by protein assembling between membranes. Understanding the mechanisms then entails determining the assembling pathway of the involved proteins. Because the intermediates are by nature transient and located in the intermembrane space, this is generally a very difficult not to say intractable problem. Here, by designing a setup with sphere/plane geometry, we have been able to freeze one transient state in which the N-terminal domains of SNARE (soluble N-ethylmaleimide–sensitive factor attachment protein receptor) proteins are assembled. A single camera frame is sufficient to obtain the complete probability of this state with the transmembrane distance. We show that it forms when membranes are 20 nm apart and stabilizes at 8 nm. This setup that fixes the intermembrane distance, and thereby the transient states, while optically probing the level of molecular assembly by Förster Resonance Energy Transfer (FRET) can be used to characterize any other transient transmembrane complexes.

Keywords: SNARE; SFA; transient state; FRET; N-terminal assembly

Domains: Cellular Biology; Biological Physics [physics.bio-ph]

Yong Jian Wang, Feng Li, Nicolas Rodriguez, Xavier Lafosse, Christine Gourier, et al.. Snapshot of sequential SNARE assembling states between membranes shows that N-terminal transient assembly initializes fusion. Proceedings of the National Academy of Sciences of the United States of America, 2016, 113 (13), pp.3533-3538. ⟨10.1073/pnas.1518935113⟩. ⟨hal-01297850⟩