Common intermediates and kinetics, but different energetics, in the assembly of SNARE proteins
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 fold...
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| Format: | Article |
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eLife Sciences Publications Ltd
2014-09-01
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| Series: | eLife |
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| Online Access: | https://elifesciences.org/articles/03348 |
| _version_ | 1811200334536638464 |
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| author | Sylvain Zorman Aleksander A Rebane Lu Ma Guangcan Yang Matthew A Molski Jeff Coleman Frederic Pincet James E Rothman Yongli Zhang |
| author_facet | Sylvain Zorman Aleksander A Rebane Lu Ma Guangcan Yang Matthew A Molski Jeff Coleman Frederic Pincet James E Rothman Yongli Zhang |
| author_sort | Sylvain Zorman |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-04-12T02:03:05Z |
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| id | doaj.art-0013692539554ac5ab8cd80a5e10e695 |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-12T02:03:05Z |
| publishDate | 2014-09-01 |
| publisher | eLife Sciences Publications Ltd |
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| series | eLife |
| spelling | doaj.art-0013692539554ac5ab8cd80a5e10e6952022-12-22T03:52:37ZengeLife Sciences Publications LtdeLife2050-084X2014-09-01310.7554/eLife.03348Common intermediates and kinetics, but different energetics, in the assembly of SNARE proteinsSylvain Zorman0Aleksander A Rebane1Lu Ma2Guangcan Yang3Matthew A Molski4Jeff Coleman5Frederic Pincet6James E Rothman7Yongli Zhang8Department of Cell Biology, Yale University School of Medicine, New Haven, United States; Nanobiology Institute, Yale University, West Haven, United StatesDepartment of Physics, Yale University, New Haven, United States; Integrated Graduate Program in Physical and Engineering Biology, Yale University, New Haven, United StatesDepartment of Cell Biology, Yale University School of Medicine, New Haven, United StatesDepartment of Cell Biology, Yale University School of Medicine, New Haven, United StatesDepartment of Cell Biology, Yale University School of Medicine, New Haven, United States; Nanobiology Institute, Yale University, West Haven, United StatesDepartment of Cell Biology, Yale University School of Medicine, New Haven, United States; Nanobiology Institute, Yale University, West Haven, United StatesDepartment of Cell Biology, Yale University School of Medicine, New Haven, United States; Nanobiology Institute, Yale University, West Haven, United States; Laboratoire de Physique Statistique, UMR CNRS 8550 Associée aux Universités Paris 6 et Paris 7, Ecole Normale Supérieure, Paris, FranceDepartment of Cell Biology, Yale University School of Medicine, New Haven, United States; Nanobiology Institute, Yale University, West Haven, United StatesDepartment of Cell Biology, Yale University School of Medicine, New Haven, United States; Nanobiology Institute, Yale University, West Haven, United StatesSoluble 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.https://elifesciences.org/articles/03348SNAREoptical tweezerprotein foldingmembrane fusionSNARE assemblyenergy landscape |
| spellingShingle | Sylvain Zorman Aleksander A Rebane Lu Ma Guangcan Yang Matthew A Molski Jeff Coleman Frederic Pincet James E Rothman Yongli Zhang Common intermediates and kinetics, but different energetics, in the assembly of SNARE proteins eLife SNARE optical tweezer protein folding membrane fusion SNARE assembly energy landscape |
| title | Common intermediates and kinetics, but different energetics, in the assembly of SNARE proteins |
| title_full | Common intermediates and kinetics, but different energetics, in the assembly of SNARE proteins |
| title_fullStr | Common intermediates and kinetics, but different energetics, in the assembly of SNARE proteins |
| title_full_unstemmed | Common intermediates and kinetics, but different energetics, in the assembly of SNARE proteins |
| title_short | Common intermediates and kinetics, but different energetics, in the assembly of SNARE proteins |
| title_sort | common intermediates and kinetics but different energetics in the assembly of snare proteins |
| topic | SNARE optical tweezer protein folding membrane fusion SNARE assembly energy landscape |
| url | https://elifesciences.org/articles/03348 |
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