Structural characterization of full-length NSF and 20S particles

Lei Fu Chang; Song Chen; Cui Cui Liu; Xijiang Pan; Jiansen Jiang; Xiao Chen Bai; Xin Xie; Hong Wei Wang; Sen Fang Sui

doi:10.1038/nsmb.2237

Structural characterization of full-length NSF and 20S particles

Lei Fu Chang, Song Chen, Cui Cui Liu, Xijiang Pan, Jiansen Jiang, Xiao Chen Bai, Xin Xie, Hong Wei Wang, Sen Fang Sui

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

44 Scopus citations

Abstract

The 20S particle, which is composed of the N-ethylmaleimide-sensitive factor (NSF), soluble NSF attachment proteins (SNAPs) and the SNAP receptor (SNARE) complex, has an essential role in intracellular vesicle fusion events. Using single-particle cryo-EM and negative stain EM, we reconstructed four related three-dimensional structures: Chinese hamster NSF hexamer in the ATPγS, ADP-AlFx and ADP states, and the 20S particle. These structures reveal a parallel arrangement between the D1 and D2 domains of the hexameric NSF and characterize the nucleotide-dependent conformational changes in NSF. The structure of the 20S particle shows that it holds the SNARE complex at two interaction interfaces around the C terminus and N-terminal half of the SNARE complex, respectively. These findings provide insight into the molecular mechanism underlying disassembly of the SNARE complex by NSF.

Original language	English (US)
Pages (from-to)	268-275
Number of pages	8
Journal	Nature Structural and Molecular Biology
Volume	19
Issue number	3
DOIs	https://doi.org/10.1038/nsmb.2237
State	Published - Mar 2012

ASJC Scopus subject areas

Structural Biology
Molecular Biology

Access to Document

10.1038/nsmb.2237

Cite this

@article{5b929380a9c94db8a1536fc59aa2338d,

title = "Structural characterization of full-length NSF and 20S particles",

abstract = "The 20S particle, which is composed of the N-ethylmaleimide-sensitive factor (NSF), soluble NSF attachment proteins (SNAPs) and the SNAP receptor (SNARE) complex, has an essential role in intracellular vesicle fusion events. Using single-particle cryo-EM and negative stain EM, we reconstructed four related three-dimensional structures: Chinese hamster NSF hexamer in the ATPγS, ADP-AlFx and ADP states, and the 20S particle. These structures reveal a parallel arrangement between the D1 and D2 domains of the hexameric NSF and characterize the nucleotide-dependent conformational changes in NSF. The structure of the 20S particle shows that it holds the SNARE complex at two interaction interfaces around the C terminus and N-terminal half of the SNARE complex, respectively. These findings provide insight into the molecular mechanism underlying disassembly of the SNARE complex by NSF.",

author = "Chang, {Lei Fu} and Song Chen and Liu, {Cui Cui} and Xijiang Pan and Jiansen Jiang and Bai, {Xiao Chen} and Xin Xie and Wang, {Hong Wei} and Sui, {Sen Fang}",

note = "Funding Information: We are grateful to E.R. Chapman (University of Wisconsin, Madison), J. Rizo (University of Texas Southwestern Medical Center) for providing plasmids. We thank F. Zhang, N. Gao, J.-W. Wu, N. Yan, J. Wang and L. Cheng for helpful discussions. We also thank J. Lei for setting up a semiautomated data collection program and Y. Wu for providing electron microscope for our use at the early stage of this work. This work was supported by the National Basic Research Program of China (2011CB910500/2010CB833706/2010CB912400) and the National Natural Science Foundation of China (30830028).",

year = "2012",

month = mar,

doi = "10.1038/nsmb.2237",

language = "English (US)",

volume = "19",

pages = "268--275",

journal = "Nature Structural and Molecular Biology",

issn = "1545-9993",

publisher = "Nature Publishing Group",

number = "3",

}

TY - JOUR

T1 - Structural characterization of full-length NSF and 20S particles

AU - Chang, Lei Fu

AU - Chen, Song

AU - Liu, Cui Cui

AU - Pan, Xijiang

AU - Jiang, Jiansen

AU - Bai, Xiao Chen

AU - Xie, Xin

AU - Wang, Hong Wei

AU - Sui, Sen Fang

N1 - Funding Information: We are grateful to E.R. Chapman (University of Wisconsin, Madison), J. Rizo (University of Texas Southwestern Medical Center) for providing plasmids. We thank F. Zhang, N. Gao, J.-W. Wu, N. Yan, J. Wang and L. Cheng for helpful discussions. We also thank J. Lei for setting up a semiautomated data collection program and Y. Wu for providing electron microscope for our use at the early stage of this work. This work was supported by the National Basic Research Program of China (2011CB910500/2010CB833706/2010CB912400) and the National Natural Science Foundation of China (30830028).

PY - 2012/3

Y1 - 2012/3

N2 - The 20S particle, which is composed of the N-ethylmaleimide-sensitive factor (NSF), soluble NSF attachment proteins (SNAPs) and the SNAP receptor (SNARE) complex, has an essential role in intracellular vesicle fusion events. Using single-particle cryo-EM and negative stain EM, we reconstructed four related three-dimensional structures: Chinese hamster NSF hexamer in the ATPγS, ADP-AlFx and ADP states, and the 20S particle. These structures reveal a parallel arrangement between the D1 and D2 domains of the hexameric NSF and characterize the nucleotide-dependent conformational changes in NSF. The structure of the 20S particle shows that it holds the SNARE complex at two interaction interfaces around the C terminus and N-terminal half of the SNARE complex, respectively. These findings provide insight into the molecular mechanism underlying disassembly of the SNARE complex by NSF.

AB - The 20S particle, which is composed of the N-ethylmaleimide-sensitive factor (NSF), soluble NSF attachment proteins (SNAPs) and the SNAP receptor (SNARE) complex, has an essential role in intracellular vesicle fusion events. Using single-particle cryo-EM and negative stain EM, we reconstructed four related three-dimensional structures: Chinese hamster NSF hexamer in the ATPγS, ADP-AlFx and ADP states, and the 20S particle. These structures reveal a parallel arrangement between the D1 and D2 domains of the hexameric NSF and characterize the nucleotide-dependent conformational changes in NSF. The structure of the 20S particle shows that it holds the SNARE complex at two interaction interfaces around the C terminus and N-terminal half of the SNARE complex, respectively. These findings provide insight into the molecular mechanism underlying disassembly of the SNARE complex by NSF.

UR - http://www.scopus.com/inward/record.url?scp=84862803840&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84862803840&partnerID=8YFLogxK

U2 - 10.1038/nsmb.2237

DO - 10.1038/nsmb.2237

M3 - Article

C2 - 22307055

AN - SCOPUS:84862803840

SN - 1545-9993

VL - 19

SP - 268

EP - 275

JO - Nature Structural and Molecular Biology

JF - Nature Structural and Molecular Biology

IS - 3

ER -

Structural characterization of full-length NSF and 20S particles

Abstract

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Cite this