TY - JOUR
T1 - Common intermediates and kinetics, but different energetics, in the assembly of SNARE proteins
AU - Zorman, Sylvain
AU - Rebane, Aleksander A.
AU - Ma, Lu
AU - Yang, Guangcan
AU - Molski, Matthew A.
AU - Coleman, Jeff
AU - Pincet, Frederic
AU - Rothman, James E.
AU - Zhang, Yongli
N1 - Funding Information:
Aleksander A Rebane, the second author, performed exemplary work in accomplishing the complex data analyses herein reported. We thank Ying Gao for technical assistance and Jingshi Shen, Daniel Kummel, and Xinming Zhang for reading the manuscript. This work was supported by the NIH grants GM093341 to YZ and DK027044 to JER.
Publisher Copyright:
© Zorman et al.
PY - 2014/9/1
Y1 - 2014/9/1
N2 - Soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs) are evolutionarily conserved machines that couple their folding/assembly to membrane fusion. However, it is unclear how these processes are regulated and function. To determine these mechanisms, we characterized the folding energy and kinetics of four representative SNARE complexes at a single-molecule level using high-resolution optical tweezers. We found that all SNARE complexes assemble by the same step-wise zippering mechanism: slow N-terminal domain (NTD) association, a pause in a force-dependent half-zippered intermediate, and fast C-terminal domain (CTD) zippering. The energy release from CTD zippering differs for yeast (13 kBT) and neuronal SNARE complexes (27 kBT), and is concentrated at the C-terminal part of CTD zippering. Thus, SNARE complexes share a conserved zippering pathway and polarized energy release to efficiently drive membrane fusion, but generate different amounts of zippering energy to regulate fusion kinetics.
AB - Soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs) are evolutionarily conserved machines that couple their folding/assembly to membrane fusion. However, it is unclear how these processes are regulated and function. To determine these mechanisms, we characterized the folding energy and kinetics of four representative SNARE complexes at a single-molecule level using high-resolution optical tweezers. We found that all SNARE complexes assemble by the same step-wise zippering mechanism: slow N-terminal domain (NTD) association, a pause in a force-dependent half-zippered intermediate, and fast C-terminal domain (CTD) zippering. The energy release from CTD zippering differs for yeast (13 kBT) and neuronal SNARE complexes (27 kBT), and is concentrated at the C-terminal part of CTD zippering. Thus, SNARE complexes share a conserved zippering pathway and polarized energy release to efficiently drive membrane fusion, but generate different amounts of zippering energy to regulate fusion kinetics.
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U2 - 10.7554/eLife.03348.001
DO - 10.7554/eLife.03348.001
M3 - Article
C2 - 25180101
AN - SCOPUS:84995679706
SN - 2050-084X
VL - 3
JO - eLife
JF - eLife
M1 - e03348
ER -