Single-molecule two- and three-colour FRET studies reveal a transition state in SNARE disassembly by NSF
Abstract SNARE (soluble N-ethylmaleimide sensitive factor attachment protein receptor) proteins are the minimal machinery required for vesicle fusion in eukaryotes. Formation of a highly stable four-helix bundle consisting of SNARE motif of these proteins, drives vesicle/membrane fusion involved in...
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| Format: | Article |
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Nature Portfolio
2025-01-01
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| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-024-55531-0 |
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| author | Sudheer K. Cheppali Chang Li Wenjing Xing Ruirui Sun Mengyi Yang Yi Xue Si-Yao Lu Jun Yao Shan Sun Chunlai Chen Sen-Fang Sui |
| author_facet | Sudheer K. Cheppali Chang Li Wenjing Xing Ruirui Sun Mengyi Yang Yi Xue Si-Yao Lu Jun Yao Shan Sun Chunlai Chen Sen-Fang Sui |
| author_sort | Sudheer K. Cheppali |
| collection | DOAJ |
| description | Abstract SNARE (soluble N-ethylmaleimide sensitive factor attachment protein receptor) proteins are the minimal machinery required for vesicle fusion in eukaryotes. Formation of a highly stable four-helix bundle consisting of SNARE motif of these proteins, drives vesicle/membrane fusion involved in several physiological processes such as neurotransmission. Recycling/disassembly of the protein machinery involved in membrane fusion is essential and is facilitated by an AAA+ ATPase, N-ethylmaleimide sensitive factor (NSF) in the presence of an adapter protein, α-SNAP. Here we use single-molecule fluorescence spectroscopy approaches to elucidate the chain of events that occur during the disassembly of SNARE complex by NSF. Our observations indicate two major pathways leading to the sequential disassembly of the SNARE complex: one where a syntaxin separated intermediate state is observed before syntaxin disassembles first, and a second where Vamp disassembles from the other proteins first. These studies uncover two parallel sequential pathways for the SNARE disassembly by NSF along with a syntaxin separated intermediate that couldn’t be observed otherwise. |
| format | Article |
| id | doaj-art-923e4970558c4218b193e375519a357e |
| institution | Kabale University |
| issn | 2041-1723 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj-art-923e4970558c4218b193e375519a357e2025-01-05T12:39:23ZengNature PortfolioNature Communications2041-17232025-01-0116111010.1038/s41467-024-55531-0Single-molecule two- and three-colour FRET studies reveal a transition state in SNARE disassembly by NSFSudheer K. Cheppali0Chang Li1Wenjing Xing2Ruirui Sun3Mengyi Yang4Yi Xue5Si-Yao Lu6Jun Yao7Shan Sun8Chunlai Chen9Sen-Fang Sui10State Key Laboratory of Membrane Biology, Beijing Frontier Research Center of Biological Structure, Beijing Advanced Innovation Center for Structural Biology, School of Life Sciences, Tsinghua UniversityState Key Laboratory of Membrane Biology, Beijing Frontier Research Center of Biological Structure, Beijing Advanced Innovation Center for Structural Biology, School of Life Sciences, Tsinghua UniversityState Key Laboratory of Membrane Biology, Beijing Frontier Research Center of Biological Structure, Beijing Advanced Innovation Center for Structural Biology, School of Life Sciences, Tsinghua UniversityState Key Laboratory of Membrane Biology, Beijing Frontier Research Center of Biological Structure, Beijing Advanced Innovation Center for Structural Biology, School of Life Sciences, Tsinghua UniversityState Key Laboratory of Membrane Biology, Beijing Frontier Research Center of Biological Structure, Beijing Advanced Innovation Center for Structural Biology, School of Life Sciences, Tsinghua UniversitySchool of Life Sciences, Tsinghua-Peking Joint Center for Life Sciences, Beijing Advanced Innovation Center for Structural Biology, Beijing Frontier Research Center of Biological Structure, Tsinghua UniversityState Key Laboratory of Membrane Biology, Tsinghua-Peking Joint Center for Life Sciences, IDG/McGovern Institute for Brain Research, School of Life Sciences, Tsinghua UniversityState Key Laboratory of Membrane Biology, Tsinghua-Peking Joint Center for Life Sciences, IDG/McGovern Institute for Brain Research, School of Life Sciences, Tsinghua UniversityState Key Laboratory of Membrane Biology, Beijing Frontier Research Center of Biological Structure, Beijing Advanced Innovation Center for Structural Biology, School of Life Sciences, Tsinghua UniversityState Key Laboratory of Membrane Biology, Beijing Frontier Research Center of Biological Structure, Beijing Advanced Innovation Center for Structural Biology, School of Life Sciences, Tsinghua UniversityState Key Laboratory of Membrane Biology, Beijing Frontier Research Center of Biological Structure, Beijing Advanced Innovation Center for Structural Biology, School of Life Sciences, Tsinghua UniversityAbstract SNARE (soluble N-ethylmaleimide sensitive factor attachment protein receptor) proteins are the minimal machinery required for vesicle fusion in eukaryotes. Formation of a highly stable four-helix bundle consisting of SNARE motif of these proteins, drives vesicle/membrane fusion involved in several physiological processes such as neurotransmission. Recycling/disassembly of the protein machinery involved in membrane fusion is essential and is facilitated by an AAA+ ATPase, N-ethylmaleimide sensitive factor (NSF) in the presence of an adapter protein, α-SNAP. Here we use single-molecule fluorescence spectroscopy approaches to elucidate the chain of events that occur during the disassembly of SNARE complex by NSF. Our observations indicate two major pathways leading to the sequential disassembly of the SNARE complex: one where a syntaxin separated intermediate state is observed before syntaxin disassembles first, and a second where Vamp disassembles from the other proteins first. These studies uncover two parallel sequential pathways for the SNARE disassembly by NSF along with a syntaxin separated intermediate that couldn’t be observed otherwise.https://doi.org/10.1038/s41467-024-55531-0 |
| spellingShingle | Sudheer K. Cheppali Chang Li Wenjing Xing Ruirui Sun Mengyi Yang Yi Xue Si-Yao Lu Jun Yao Shan Sun Chunlai Chen Sen-Fang Sui Single-molecule two- and three-colour FRET studies reveal a transition state in SNARE disassembly by NSF Nature Communications |
| title | Single-molecule two- and three-colour FRET studies reveal a transition state in SNARE disassembly by NSF |
| title_full | Single-molecule two- and three-colour FRET studies reveal a transition state in SNARE disassembly by NSF |
| title_fullStr | Single-molecule two- and three-colour FRET studies reveal a transition state in SNARE disassembly by NSF |
| title_full_unstemmed | Single-molecule two- and three-colour FRET studies reveal a transition state in SNARE disassembly by NSF |
| title_short | Single-molecule two- and three-colour FRET studies reveal a transition state in SNARE disassembly by NSF |
| title_sort | single molecule two and three colour fret studies reveal a transition state in snare disassembly by nsf |
| url | https://doi.org/10.1038/s41467-024-55531-0 |
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