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Title: Architecture of the synaptotagmin–SNARE machinery for neuronal exocytosis

Abstract

Synaptotagmin-1 and neuronal SNARE proteins have central roles in evoked synchronous neurotransmitter release; however, it is unknown how they cooperate to trigger synaptic vesicle fusion. We report atomic-resolution crystal structures of Ca 2+- and Mg 2+-bound complexes between synaptotagmin-1 and the neuronal SNARE complex, one of which was determined with diffraction data from an X-ray free-electron laser, leading to an atomic-resolution structure with accurate rotamer assignments for many side chains. The structures reveal several interfaces, including a large, specific, Ca 2+-independent and conserved interface. Tests of this interface by mutagenesis suggest that it is essential for Ca 2+-triggered neurotransmitter release in mouse hippocampal neuronal synapses and for Ca 2+-triggered vesicle fusion in a reconstituted system. Lastly, we propose that this interface forms before Ca 2+ triggering, moves en bloc as Ca 2+ influx promotes the interactions between synaptotagmin-1 and the plasma membrane, and consequently remodels the membrane to promote fusion, possibly in conjunction with other interfaces.

Authors:
 [1];  [1];  [2];  [1];  [1];  [1];  [1];  [3];  [3];  [4];  [4];  [4];  [4];  [4];  [1];  [1];  [5];  [1];  [2];  [1]
  1. Stanford Univ., CA (United States). Howard Hughes Medical Institute; Stanford Univ., CA (United States). Departments of Neurology and Neurological Sciences, Photon Science, and Structural Biology
  2. Stanford Univ., CA (United States). Howard Hughes Medical Institute
  3. Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
  4. SLAC National Accelerator Lab., Stanford, CA (United States)
  5. Stanford Univ., CA (United States). Departments of Structural Biology, Molecular and Cellular Physiology, and Photon Science
Publication Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22); USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23)
Contributing Org.:
National Institutes of Health (NIH)
OSTI Identifier:
1221448
Alternate Identifier(s):
OSTI ID: 1512250
Grant/Contract Number:  
AC02-76SF00515; P41GM103393; AC02-05CH11231
Resource Type:
Accepted Manuscript
Journal Name:
Nature (London)
Additional Journal Information:
Journal Name: Nature (London); Journal Volume: 525; Journal Issue: 7567; Journal ID: ISSN 0028-0836
Publisher:
Nature Publishing Group
Country of Publication:
United States
Language:
ENGLISH
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Zhou, Qiangjun, Lai, Ying, Bacaj, Taulant, Zhao, Minglei, Lyubimov, Artem Y., Uervirojnangkoorn, Monarin, Zeldin, Oliver B., Brewster, Aaron S., Sauter, Nicholas K., Cohen, Aina E., Soltis, S. Michael, Alonso-Mori, Roberto, Chollet, Matthieu, Lemke, Henrik T., Pfuetzner, Richard A., Choi, Ucheor B., Weis, William I., Diao, Jiajie, Südhof, Thomas C., and Brunger, Axel T. Architecture of the synaptotagmin–SNARE machinery for neuronal exocytosis. United States: N. p., 2015. Web. doi:10.1038/nature14975.
Zhou, Qiangjun, Lai, Ying, Bacaj, Taulant, Zhao, Minglei, Lyubimov, Artem Y., Uervirojnangkoorn, Monarin, Zeldin, Oliver B., Brewster, Aaron S., Sauter, Nicholas K., Cohen, Aina E., Soltis, S. Michael, Alonso-Mori, Roberto, Chollet, Matthieu, Lemke, Henrik T., Pfuetzner, Richard A., Choi, Ucheor B., Weis, William I., Diao, Jiajie, Südhof, Thomas C., & Brunger, Axel T. Architecture of the synaptotagmin–SNARE machinery for neuronal exocytosis. United States. doi:10.1038/nature14975.
Zhou, Qiangjun, Lai, Ying, Bacaj, Taulant, Zhao, Minglei, Lyubimov, Artem Y., Uervirojnangkoorn, Monarin, Zeldin, Oliver B., Brewster, Aaron S., Sauter, Nicholas K., Cohen, Aina E., Soltis, S. Michael, Alonso-Mori, Roberto, Chollet, Matthieu, Lemke, Henrik T., Pfuetzner, Richard A., Choi, Ucheor B., Weis, William I., Diao, Jiajie, Südhof, Thomas C., and Brunger, Axel T. Mon . "Architecture of the synaptotagmin–SNARE machinery for neuronal exocytosis". United States. doi:10.1038/nature14975. https://www.osti.gov/servlets/purl/1221448.
@article{osti_1221448,
title = {Architecture of the synaptotagmin–SNARE machinery for neuronal exocytosis},
author = {Zhou, Qiangjun and Lai, Ying and Bacaj, Taulant and Zhao, Minglei and Lyubimov, Artem Y. and Uervirojnangkoorn, Monarin and Zeldin, Oliver B. and Brewster, Aaron S. and Sauter, Nicholas K. and Cohen, Aina E. and Soltis, S. Michael and Alonso-Mori, Roberto and Chollet, Matthieu and Lemke, Henrik T. and Pfuetzner, Richard A. and Choi, Ucheor B. and Weis, William I. and Diao, Jiajie and Südhof, Thomas C. and Brunger, Axel T.},
abstractNote = {Synaptotagmin-1 and neuronal SNARE proteins have central roles in evoked synchronous neurotransmitter release; however, it is unknown how they cooperate to trigger synaptic vesicle fusion. We report atomic-resolution crystal structures of Ca2+- and Mg2+-bound complexes between synaptotagmin-1 and the neuronal SNARE complex, one of which was determined with diffraction data from an X-ray free-electron laser, leading to an atomic-resolution structure with accurate rotamer assignments for many side chains. The structures reveal several interfaces, including a large, specific, Ca2+-independent and conserved interface. Tests of this interface by mutagenesis suggest that it is essential for Ca2+-triggered neurotransmitter release in mouse hippocampal neuronal synapses and for Ca2+-triggered vesicle fusion in a reconstituted system. Lastly, we propose that this interface forms before Ca2+ triggering, moves en bloc as Ca2+ influx promotes the interactions between synaptotagmin-1 and the plasma membrane, and consequently remodels the membrane to promote fusion, possibly in conjunction with other interfaces.},
doi = {10.1038/nature14975},
journal = {Nature (London)},
number = 7567,
volume = 525,
place = {United States},
year = {2015},
month = {8}
}

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