title: Spring-loaded unraveling of a single SNARE complex by NSF in one round of ATP turnover.
abstract: During intracellular membrane trafficking, N-ethylmaleimide-sensitive factor (NSF) and alpha-soluble NSF attachment protein (α-SNAP) disassemble the soluble NSF attachment protein receptor (SNARE) complex for recycling of the SNARE proteins. The molecular mechanism by which NSF disassembles the SNARE complex is largely unknown. Using single-molecule fluorescence spectroscopy and magnetic tweezers, we found that NSF disassembled a single SNARE complex in only one round of adenosine triphosphate (ATP) turnover. Upon ATP cleavage, the NSF hexamer developed internal tension with dissociation of phosphate ions. After latent time measuring tens of seconds, NSF released the built-up tension in a burst within 20 milliseconds, resulting in disassembly followed by immediate release of the SNARE proteins. Thus, NSF appears to use a "spring-loaded" mechanism to couple ATP hydrolysis and unfolding of substrate proteins.
authors: National Creative Research Initiative Center for Single-Molecule Systems Biology, Korea Advanced Institute of Science and Technology (KAIST), Daejeon 305-701, South Korea. Department of Physics, KAIST, Daejeon 305-701, South Korea.
authors: Je-Kyung
authors: JK
authors: Ryu
authors: National Creative Research Initiative Center for Single-Molecule Systems Biology, Korea Advanced Institute of Science and Technology (KAIST), Daejeon 305-701, South Korea. Department of Physics, KAIST, Daejeon 305-701, South Korea.
authors: Duyoung
authors: D
authors: Min
authors: National Creative Research Initiative Center for Single-Molecule Systems Biology, Korea Advanced Institute of Science and Technology (KAIST), Daejeon 305-701, South Korea. Department of Physics, KAIST, Daejeon 305-701, South Korea.
authors: Sang-Hyun
authors: SH
authors: Rah
authors: Graduate School of Medical Science and Engineering, KAIST, Daejeon 305-701, South Korea.
authors: Soo Jin
authors: SJ
authors: Kim
authors: Department of Neurobiology, Max-Planck-Institute for Biophysical Chemistry, 37077 Göttingen, Germany.
authors: Yongsoo
authors: Y
authors: Park
authors: Graduate School of Medical Science and Engineering, KAIST, Daejeon 305-701, South Korea.
authors: Haesoo
authors: H
authors: Kim
authors: Korea Institute for Advanced Study, Seoul 130-722, South Korea.
authors: Changbong
authors: C
authors: Hyeon
authors: Graduate School of Medical Science and Engineering, KAIST, Daejeon 305-701, South Korea.
authors: Ho Min
authors: HM
authors: Kim
authors: Department of Neurobiology, Max-Planck-Institute for Biophysical Chemistry, 37077 Göttingen, Germany. rjahn@gwdg.de tyyoon@kaist.ac.kr.
authors: Reinhard
authors: R
authors: Jahn
authors: National Creative Research Initiative Center for Single-Molecule Systems Biology, Korea Advanced Institute of Science and Technology (KAIST), Daejeon 305-701, South Korea. Department of Physics, KAIST, Daejeon 305-701, South Korea. rjahn@gwdg.de tyyoon@kaist.ac.kr.
authors: Tae-Young
authors: TY
authors: Yoon
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism*; D000818:Animals; D002417:Cattle; D006224:Cricetinae; D031541:Fluorescence Resonance Energy Transfer; D006868:Hydrolysis; D052066:N-Ethylmaleimide-Sensitive Proteins / Q000378:metabolism*; D051381:Rats; D050600:SNARE Proteins / Q000378:metabolism*; D052067:Soluble N-Ethylmaleimide-Sensitive Factor Attachment Proteins / Q000378:metabolism*; D013050:Spectrometry, Fluorescence
chemical_list: D050600:SNARE Proteins; D052067:Soluble N-Ethylmaleimide-Sensitive Factor Attachment Proteins; D000255:Adenosine Triphosphate; D052066:N-Ethylmaleimide-Sensitive Proteins