Synaptic proteins promote calcium-triggered fast transition from point contact to full fusion
The molecular underpinnings of synaptic vesicle fusion for fast neurotransmitter release are still unclear. Here, we used a single vesicle–vesicle system with reconstituted SNARE and synaptotagmin-1 proteoliposomes to decipher the temporal sequence of membrane states upon Ca2+-injection at 250–500 μ...
| Main Authors: | , , , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
eLife Sciences Publications Ltd
2012-12-01
|
| Series: | eLife |
| Subjects: | |
| Online Access: | https://elifesciences.org/articles/00109 |
| _version_ | 1811200039990591488 |
|---|---|
| author | Jiajie Diao Patricia Grob Daniel J Cipriano Minjoung Kyoung Yunxiang Zhang Sachi Shah Amie Nguyen Mark Padolina Ankita Srivastava Marija Vrljic Ankita Shah Eva Nogales Steven Chu Axel T Brunger |
| author_facet | Jiajie Diao Patricia Grob Daniel J Cipriano Minjoung Kyoung Yunxiang Zhang Sachi Shah Amie Nguyen Mark Padolina Ankita Srivastava Marija Vrljic Ankita Shah Eva Nogales Steven Chu Axel T Brunger |
| author_sort | Jiajie Diao |
| collection | DOAJ |
| description | The molecular underpinnings of synaptic vesicle fusion for fast neurotransmitter release are still unclear. Here, we used a single vesicle–vesicle system with reconstituted SNARE and synaptotagmin-1 proteoliposomes to decipher the temporal sequence of membrane states upon Ca2+-injection at 250–500 μM on a 100-ms timescale. Furthermore, detailed membrane morphologies were imaged with cryo-electron microscopy before and after Ca2+-injection. We discovered a heterogeneous network of immediate and delayed fusion pathways. Remarkably, all instances of Ca2+-triggered immediate fusion started from a membrane–membrane point-contact and proceeded to complete fusion without discernible hemifusion intermediates. In contrast, pathways that involved a stable hemifusion diaphragm only resulted in fusion after many seconds, if at all. When complexin was included, the Ca2+-triggered fusion network shifted towards the immediate pathway, effectively synchronizing fusion, especially at lower Ca2+-concentration. Synaptic proteins may have evolved to select this immediate pathway out of a heterogeneous network of possible membrane fusion pathways. |
| first_indexed | 2024-04-12T01:58:03Z |
| format | Article |
| id | doaj.art-7439d9935d5b41ab9eb3efd1ac2fd2d9 |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-12T01:58:03Z |
| publishDate | 2012-12-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-7439d9935d5b41ab9eb3efd1ac2fd2d92022-12-22T03:52:45ZengeLife Sciences Publications LtdeLife2050-084X2012-12-01110.7554/eLife.00109Synaptic proteins promote calcium-triggered fast transition from point contact to full fusionJiajie Diao0Patricia Grob1Daniel J Cipriano2Minjoung Kyoung3Yunxiang Zhang4Sachi Shah5Amie Nguyen6Mark Padolina7Ankita Srivastava8Marija Vrljic9Ankita Shah10Eva Nogales11Steven Chu12Axel T Brunger13Departments of Molecular and Cellular Physiology, Neurology and Neurological Sciences, Structural Biology, Photon Science and Howard Hughes Medical Institute, Stanford University, Stanford, USADepartment of Molecular and Cell Biology and Howard Hughes Medical Institute, University of California at Berkeley, Berkeley, USADepartments of Molecular and Cellular Physiology, Neurology and Neurological Sciences, Structural Biology, Photon Science and Howard Hughes Medical Institute, Stanford University, Stanford, USADepartments of Molecular and Cellular Physiology, Neurology and Neurological Sciences, Structural Biology, Photon Science and Howard Hughes Medical Institute, Stanford University, Stanford, USADepartments of Molecular and Cellular Physiology, Neurology and Neurological Sciences, Structural Biology, Photon Science and Howard Hughes Medical Institute, Stanford University, Stanford, USADepartments of Molecular and Cellular Physiology, Neurology and Neurological Sciences, Structural Biology, Photon Science and Howard Hughes Medical Institute, Stanford University, Stanford, USADepartments of Molecular and Cellular Physiology, Neurology and Neurological Sciences, Structural Biology, Photon Science and Howard Hughes Medical Institute, Stanford University, Stanford, USADepartments of Molecular and Cellular Physiology, Neurology and Neurological Sciences, Structural Biology, Photon Science and Howard Hughes Medical Institute, Stanford University, Stanford, USADepartments of Molecular and Cellular Physiology, Neurology and Neurological Sciences, Structural Biology, Photon Science and Howard Hughes Medical Institute, Stanford University, Stanford, USADepartments of Molecular and Cellular Physiology, Neurology and Neurological Sciences, Structural Biology, Photon Science and Howard Hughes Medical Institute, Stanford University, Stanford, USADepartments of Molecular and Cellular Physiology, Neurology and Neurological Sciences, Structural Biology, Photon Science and Howard Hughes Medical Institute, Stanford University, Stanford, USADepartment of Molecular and Cell Biology and Howard Hughes Medical Institute, University of California at Berkeley, Berkeley, USA; Life Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, USAFormerly Lawrence Berkeley National Laboratory, and Departments of Physics and Molecular and Cell Biology, University of California at Berkeley, Berkeley, USADepartments of Molecular and Cellular Physiology, Neurology and Neurological Sciences, Structural Biology, Photon Science and Howard Hughes Medical Institute, Stanford University, Stanford, USAThe molecular underpinnings of synaptic vesicle fusion for fast neurotransmitter release are still unclear. Here, we used a single vesicle–vesicle system with reconstituted SNARE and synaptotagmin-1 proteoliposomes to decipher the temporal sequence of membrane states upon Ca2+-injection at 250–500 μM on a 100-ms timescale. Furthermore, detailed membrane morphologies were imaged with cryo-electron microscopy before and after Ca2+-injection. We discovered a heterogeneous network of immediate and delayed fusion pathways. Remarkably, all instances of Ca2+-triggered immediate fusion started from a membrane–membrane point-contact and proceeded to complete fusion without discernible hemifusion intermediates. In contrast, pathways that involved a stable hemifusion diaphragm only resulted in fusion after many seconds, if at all. When complexin was included, the Ca2+-triggered fusion network shifted towards the immediate pathway, effectively synchronizing fusion, especially at lower Ca2+-concentration. Synaptic proteins may have evolved to select this immediate pathway out of a heterogeneous network of possible membrane fusion pathways.https://elifesciences.org/articles/00109neurotransmitter releasesynaptic vesicle fusionSNAREsynaptotagmincomplexin |
| spellingShingle | Jiajie Diao Patricia Grob Daniel J Cipriano Minjoung Kyoung Yunxiang Zhang Sachi Shah Amie Nguyen Mark Padolina Ankita Srivastava Marija Vrljic Ankita Shah Eva Nogales Steven Chu Axel T Brunger Synaptic proteins promote calcium-triggered fast transition from point contact to full fusion eLife neurotransmitter release synaptic vesicle fusion SNARE synaptotagmin complexin |
| title | Synaptic proteins promote calcium-triggered fast transition from point contact to full fusion |
| title_full | Synaptic proteins promote calcium-triggered fast transition from point contact to full fusion |
| title_fullStr | Synaptic proteins promote calcium-triggered fast transition from point contact to full fusion |
| title_full_unstemmed | Synaptic proteins promote calcium-triggered fast transition from point contact to full fusion |
| title_short | Synaptic proteins promote calcium-triggered fast transition from point contact to full fusion |
| title_sort | synaptic proteins promote calcium triggered fast transition from point contact to full fusion |
| topic | neurotransmitter release synaptic vesicle fusion SNARE synaptotagmin complexin |
| url | https://elifesciences.org/articles/00109 |
| work_keys_str_mv | AT jiajiediao synapticproteinspromotecalciumtriggeredfasttransitionfrompointcontacttofullfusion AT patriciagrob synapticproteinspromotecalciumtriggeredfasttransitionfrompointcontacttofullfusion AT danieljcipriano synapticproteinspromotecalciumtriggeredfasttransitionfrompointcontacttofullfusion AT minjoungkyoung synapticproteinspromotecalciumtriggeredfasttransitionfrompointcontacttofullfusion AT yunxiangzhang synapticproteinspromotecalciumtriggeredfasttransitionfrompointcontacttofullfusion AT sachishah synapticproteinspromotecalciumtriggeredfasttransitionfrompointcontacttofullfusion AT amienguyen synapticproteinspromotecalciumtriggeredfasttransitionfrompointcontacttofullfusion AT markpadolina synapticproteinspromotecalciumtriggeredfasttransitionfrompointcontacttofullfusion AT ankitasrivastava synapticproteinspromotecalciumtriggeredfasttransitionfrompointcontacttofullfusion AT marijavrljic synapticproteinspromotecalciumtriggeredfasttransitionfrompointcontacttofullfusion AT ankitashah synapticproteinspromotecalciumtriggeredfasttransitionfrompointcontacttofullfusion AT evanogales synapticproteinspromotecalciumtriggeredfasttransitionfrompointcontacttofullfusion AT stevenchu synapticproteinspromotecalciumtriggeredfasttransitionfrompointcontacttofullfusion AT axeltbrunger synapticproteinspromotecalciumtriggeredfasttransitionfrompointcontacttofullfusion |