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Single-molecule two- and three-colour FRET studies reveal a transition state in SNARE disassembly by NSF

Author

Listed:
  • Sudheer K. Cheppali

    (Tsinghua University)

  • Chang Li

    (Tsinghua University)

  • Wenjing Xing

    (Tsinghua University)

  • Ruirui Sun

    (Tsinghua University)

  • Mengyi Yang

    (Tsinghua University)

  • Yi Xue

    (Tsinghua University)

  • Si-Yao Lu

    (Tsinghua University)

  • Jun Yao

    (Tsinghua University)

  • Shan Sun

    (Tsinghua University)

  • Chunlai Chen

    (Tsinghua University)

  • Sen-Fang Sui

    (Tsinghua University
    Southern University of Science and Technology)

Abstract

SNARE (soluble N-ethylmaleimide sensitive factor attachment protein receptor) proteins are the minimal machinery required for vesicle fusion in eukaryotes. Formation of a highly stable four-helix bundle consisting of SNARE motif of these proteins, drives vesicle/membrane fusion involved in several physiological processes such as neurotransmission. Recycling/disassembly of the protein machinery involved in membrane fusion is essential and is facilitated by an AAA+ ATPase, N-ethylmaleimide sensitive factor (NSF) in the presence of an adapter protein, α-SNAP. Here we use single-molecule fluorescence spectroscopy approaches to elucidate the chain of events that occur during the disassembly of SNARE complex by NSF. Our observations indicate two major pathways leading to the sequential disassembly of the SNARE complex: one where a syntaxin separated intermediate state is observed before syntaxin disassembles first, and a second where Vamp disassembles from the other proteins first. These studies uncover two parallel sequential pathways for the SNARE disassembly by NSF along with a syntaxin separated intermediate that couldn’t be observed otherwise.

Suggested Citation

  • Sudheer K. Cheppali & Chang Li & Wenjing Xing & Ruirui Sun & Mengyi Yang & Yi Xue & Si-Yao Lu & Jun Yao & Shan Sun & Chunlai Chen & Sen-Fang Sui, 2025. "Single-molecule two- and three-colour FRET studies reveal a transition state in SNARE disassembly by NSF," Nature Communications, Nature, vol. 16(1), pages 1-10, December.
    • Handle: RePEc:nat:natcom:v:16:y:2025:i:1:d:10.1038_s41467-024-55531-0
    DOI: 10.1038/s41467-024-55531-0
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