| Title: |
Nanodisc-cell fusion: control of fusion pore nucleation and lifetimes by SNARE protein transmembrane domains |
| Authors: |
Wu, Z; Auclair, SM; Bello, O; Vennekate, W; Dudzinski, NR; Krishnakumar, SS; Karatekin, E |
| Source: |
Scientific Reports , 6 , Article 27287. (2016) |
| Publisher Information: |
NATURE PUBLISHING GROUP |
| Publication Year: |
2016 |
| Collection: |
University College London: UCL Discovery |
| Subject Terms: |
Science & Technology; Multidisciplinary Sciences; Science & Technology - Other Topics; ADRENAL CHROMAFFIN CELLS; MEMBRANE-FUSION; TRANSMITTER RELEASE; TRANSIENT PORES; IN-VITRO; EXOCYTOSIS; VESICLES; PLANAR; PATCH; CLAMP |
| Description: |
The initial, nanometer-sized connection between the plasma membrane and a hormone- or neurotransmitter-filled vesicle –the fusion pore– can flicker open and closed repeatedly before dilating or resealing irreversibly. Pore dynamics determine release and vesicle recycling kinetics, but pore properties are poorly known because biochemically defined single-pore assays are lacking. We isolated single flickering pores connecting v-SNARE-reconstituted nanodiscs to cells ectopically expressing cognate, “flipped” t-SNAREs. Conductance through single, voltage-clamped fusion pores directly reported sub-millisecond pore dynamics. Pore currents fluctuated, transiently returned to baseline multiple times, and disappeared ~6 s after initial opening, as if the fusion pore fluctuated in size, flickered, and resealed. We found that interactions between v- and t-SNARE transmembrane domains (TMDs) promote, but are not essential for pore nucleation. Surprisingly, TMD modifications designed to disrupt v- and t-SNARE TMD zippering prolonged pore lifetimes dramatically. We propose that the post-fusion geometry of the proteins contribute to pore stability. |
| Document Type: |
article in journal/newspaper |
| File Description: |
text |
| Language: |
English |
| Relation: |
https://discovery.ucl.ac.uk/id/eprint/1541643/1/Bello_srep27287.pdf; https://discovery.ucl.ac.uk/id/eprint/1541643/ |
| Availability: |
https://discovery.ucl.ac.uk/id/eprint/1541643/1/Bello_srep27287.pdf; https://discovery.ucl.ac.uk/id/eprint/1541643/ |
| Rights: |
open |
| Accession Number: |
edsbas.E83D6D74 |
| Database: |
BASE |