The sec1/munc18 protein vps45 holds the qa-snare tlg2 in an open conformation

Travis J. Eisemann; Frederick Allen; Kelly Lau; Gregory R. Shimamura; Philip D. Jeffrey; Frederick M. Hughson

doi:https://doi.org/10.7554/ELIFE.60724

The sec1/munc18 protein vps45 holds the qa-snare tlg2 in an open conformation

Travis J. Eisemann, Frederick Allen, Kelly Lau, Gregory R. Shimamura, Philip D. Jeffrey, Frederick M. Hughson

Princeton University

Research output: Contribution to journal › Article › peer-review

1 Scopus citations

Abstract

Fusion of intracellular trafficking vesicles is mediated by the assembly of SNARE proteins into membrane-bridging complexes. SNARE-mediated membrane fusion requires Sec1/ Munc18-family (SM) proteins, SNARE chaperones that can function as templates to catalyze SNARE complex assembly. Paradoxically, the SM protein Munc18-1 traps the Qa-SNARE protein syntaxin-1 in an autoinhibited closed conformation. Here we present the structure of a second SM–Qa-SNARE complex, Vps45–Tlg2. Strikingly, Vps45 holds Tlg2 in an open conformation, with its SNARE motif disengaged from its Habc domain and its linker region unfolded. The domain 3a helical hairpin of Vps45 is unfurled, exposing the presumptive R-SNARE binding site to allow template complex formation. Although Tlg2 has a pronounced tendency to form homo-tetramers, Vps45 can rescue Tlg2 tetramers into stoichiometric Vps45–Tlg2 complexes. Our findings demonstrate that SM proteins can engage Qa-SNAREs using at least two different modes, one in which the SNARE is closed and one in which it is open.

Original language	English (US)
Article number	e60724
Pages (from-to)	1-18
Number of pages	18
Journal	eLife
Volume	9
DOIs	https://doi.org/10.7554/ELIFE.60724
State	Published - Aug 2020

ASJC Scopus subject areas

Biochemistry, Genetics and Molecular Biology(all)
Immunology and Microbiology(all)
Neuroscience(all)

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https://doi.org/10.7554/ELIFE.60724

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@article{8dc5ca72c5794a1fbd436b0ed266364c,

title = "The sec1/munc18 protein vps45 holds the qa-snare tlg2 in an open conformation",

abstract = "Fusion of intracellular trafficking vesicles is mediated by the assembly of SNARE proteins into membrane-bridging complexes. SNARE-mediated membrane fusion requires Sec1/ Munc18-family (SM) proteins, SNARE chaperones that can function as templates to catalyze SNARE complex assembly. Paradoxically, the SM protein Munc18-1 traps the Qa-SNARE protein syntaxin-1 in an autoinhibited closed conformation. Here we present the structure of a second SM–Qa-SNARE complex, Vps45–Tlg2. Strikingly, Vps45 holds Tlg2 in an open conformation, with its SNARE motif disengaged from its Habc domain and its linker region unfolded. The domain 3a helical hairpin of Vps45 is unfurled, exposing the presumptive R-SNARE binding site to allow template complex formation. Although Tlg2 has a pronounced tendency to form homo-tetramers, Vps45 can rescue Tlg2 tetramers into stoichiometric Vps45–Tlg2 complexes. Our findings demonstrate that SM proteins can engage Qa-SNAREs using at least two different modes, one in which the SNARE is closed and one in which it is open.",

author = "Eisemann, {Travis J.} and Frederick Allen and Kelly Lau and Shimamura, {Gregory R.} and Jeffrey, {Philip D.} and Hughson, {Frederick M.}",

note = "Funding Information: We thank Mary Munson, Jose Rizo, Venu Vandavasi, Yongli Zhang, and members of the Hughson laboratory past and present for helpful advice and discussion. The Princeton Biophysics and Macromolecular Crystallography core facilities provided essential assistance with analytical ultracentrifugation and X-ray crystallography, respectively. This work was supported by National Institutes of Health (NIH) grants T32GM007388 (GRS) and R01GM071574 (FMH). This research used the AMX and FMX beamlines of the National Synchrotron Light Source II, a United States Department of Energy (DOE) Office of Science User Facility operated for the DOE Office of Science by Brookhaven National Laboratory under Contract No. DE-SC0012704. The Center for BioMolecular Structure (CBMS) is primarily supported by the NIH, National Institute of General Medical Sciences (NIGMS) through a Center Core P30 Grant (P30GM133893), and by the DOE Office of Biological and Environmental Research (KP1605010).This work is based upon research conducted at the Cornell High Energy Synchrotron Source (CHESS), which is supported by the National Science Foundation (NSF) and the NIH/NIGMS under NSF award DMR-1829070, using the Macromolecular Diffraction at CHESS (MacCHESS) facility, which is supported by NIH/NIGMS award GM-124166. Funding Information: We thank Mary Munson, Jose Rizo, Venu Vandavasi, Yongli Zhang, and members of the Hughson laboratory past and present for helpful advice and discussion. The Princeton Biophysics and Macro-molecular Crystallography core facilities provided essential assistance with analytical ultracentrifuga-tion and X-ray crystallography, respectively. This work was supported by National Institutes of Health (NIH) grants T32GM007388 (GRS) and R01GM071574 (FMH). This research used the AMX and FMX beamlines of the National Synchrotron Light Source II, a United States Department of Energy (DOE) Office of Science User Facility operated for the DOE Office of Science by Brookhaven National Laboratory under Contract No. DE-SC0012704. The Center for BioMolecular Structure (CBMS) is primarily supported by the NIH, National Institute of General Medical Sciences (NIGMS) through a Center Core P30 Grant (P30GM133893), and by the DOE Office of Biological and Environmental Research (KP1605010).This work is based upon research conducted at the Cornell High Energy Synchrotron Source (CHESS), which is supported by the National Science Foundation (NSF) and the NIH/NIGMS under NSF award DMR-1829070, using the Macromolecular Diffraction at CHESS (MacCHESS) facility, which is supported by NIH/NIGMS award GM-124166. Publisher Copyright: {\textcopyright} Eisemann et al.",

year = "2020",

month = aug,

doi = "https://doi.org/10.7554/ELIFE.60724",

language = "English (US)",

volume = "9",

pages = "1--18",

journal = "eLife",

issn = "2050-084X",

publisher = "eLife Sciences Publications",

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T1 - The sec1/munc18 protein vps45 holds the qa-snare tlg2 in an open conformation

AU - Eisemann, Travis J.

AU - Allen, Frederick

AU - Lau, Kelly

AU - Shimamura, Gregory R.

AU - Jeffrey, Philip D.

AU - Hughson, Frederick M.

N1 - Funding Information: We thank Mary Munson, Jose Rizo, Venu Vandavasi, Yongli Zhang, and members of the Hughson laboratory past and present for helpful advice and discussion. The Princeton Biophysics and Macromolecular Crystallography core facilities provided essential assistance with analytical ultracentrifugation and X-ray crystallography, respectively. This work was supported by National Institutes of Health (NIH) grants T32GM007388 (GRS) and R01GM071574 (FMH). This research used the AMX and FMX beamlines of the National Synchrotron Light Source II, a United States Department of Energy (DOE) Office of Science User Facility operated for the DOE Office of Science by Brookhaven National Laboratory under Contract No. DE-SC0012704. The Center for BioMolecular Structure (CBMS) is primarily supported by the NIH, National Institute of General Medical Sciences (NIGMS) through a Center Core P30 Grant (P30GM133893), and by the DOE Office of Biological and Environmental Research (KP1605010).This work is based upon research conducted at the Cornell High Energy Synchrotron Source (CHESS), which is supported by the National Science Foundation (NSF) and the NIH/NIGMS under NSF award DMR-1829070, using the Macromolecular Diffraction at CHESS (MacCHESS) facility, which is supported by NIH/NIGMS award GM-124166. Funding Information: We thank Mary Munson, Jose Rizo, Venu Vandavasi, Yongli Zhang, and members of the Hughson laboratory past and present for helpful advice and discussion. The Princeton Biophysics and Macro-molecular Crystallography core facilities provided essential assistance with analytical ultracentrifuga-tion and X-ray crystallography, respectively. This work was supported by National Institutes of Health (NIH) grants T32GM007388 (GRS) and R01GM071574 (FMH). This research used the AMX and FMX beamlines of the National Synchrotron Light Source II, a United States Department of Energy (DOE) Office of Science User Facility operated for the DOE Office of Science by Brookhaven National Laboratory under Contract No. DE-SC0012704. The Center for BioMolecular Structure (CBMS) is primarily supported by the NIH, National Institute of General Medical Sciences (NIGMS) through a Center Core P30 Grant (P30GM133893), and by the DOE Office of Biological and Environmental Research (KP1605010).This work is based upon research conducted at the Cornell High Energy Synchrotron Source (CHESS), which is supported by the National Science Foundation (NSF) and the NIH/NIGMS under NSF award DMR-1829070, using the Macromolecular Diffraction at CHESS (MacCHESS) facility, which is supported by NIH/NIGMS award GM-124166. Publisher Copyright: © Eisemann et al.

PY - 2020/8

Y1 - 2020/8

N2 - Fusion of intracellular trafficking vesicles is mediated by the assembly of SNARE proteins into membrane-bridging complexes. SNARE-mediated membrane fusion requires Sec1/ Munc18-family (SM) proteins, SNARE chaperones that can function as templates to catalyze SNARE complex assembly. Paradoxically, the SM protein Munc18-1 traps the Qa-SNARE protein syntaxin-1 in an autoinhibited closed conformation. Here we present the structure of a second SM–Qa-SNARE complex, Vps45–Tlg2. Strikingly, Vps45 holds Tlg2 in an open conformation, with its SNARE motif disengaged from its Habc domain and its linker region unfolded. The domain 3a helical hairpin of Vps45 is unfurled, exposing the presumptive R-SNARE binding site to allow template complex formation. Although Tlg2 has a pronounced tendency to form homo-tetramers, Vps45 can rescue Tlg2 tetramers into stoichiometric Vps45–Tlg2 complexes. Our findings demonstrate that SM proteins can engage Qa-SNAREs using at least two different modes, one in which the SNARE is closed and one in which it is open.

AB - Fusion of intracellular trafficking vesicles is mediated by the assembly of SNARE proteins into membrane-bridging complexes. SNARE-mediated membrane fusion requires Sec1/ Munc18-family (SM) proteins, SNARE chaperones that can function as templates to catalyze SNARE complex assembly. Paradoxically, the SM protein Munc18-1 traps the Qa-SNARE protein syntaxin-1 in an autoinhibited closed conformation. Here we present the structure of a second SM–Qa-SNARE complex, Vps45–Tlg2. Strikingly, Vps45 holds Tlg2 in an open conformation, with its SNARE motif disengaged from its Habc domain and its linker region unfolded. The domain 3a helical hairpin of Vps45 is unfurled, exposing the presumptive R-SNARE binding site to allow template complex formation. Although Tlg2 has a pronounced tendency to form homo-tetramers, Vps45 can rescue Tlg2 tetramers into stoichiometric Vps45–Tlg2 complexes. Our findings demonstrate that SM proteins can engage Qa-SNAREs using at least two different modes, one in which the SNARE is closed and one in which it is open.

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The sec1/munc18 protein vps45 holds the qa-snare tlg2 in an open conformation

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