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Structural Dynamics of SNARE Complex Assembly in the Ribbon Synapses Observed by smFRET

  • Alexa L. Andrzejewski
  • , Joshua Ferrar
  • , Marla Kratzer-Charron
  • , Mark E. Bowen
  • , Ucheor B. Choi
  • Quantum-Si
  • West Virginia University

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

Abstract

Single-molecule fluorescence resonance energy transfer (smFRET) is a powerful technique for studying the structural dynamics of protein molecules or detecting interactions between protein molecules in real time. Due to the high sensitivity in spatial and temporal resolution, smFRET can decipher sub-populations within heterogeneous native state conformations, which are generally lost in traditional measurements due to ensemble averaging. In addition, the single-molecule reconstitution allows protein molecules to be observed for an extensive period of time and can recapitulate the geometry of the cellular environment to retain biological function. Here we provide a detailed method of using smFRET to monitor the conformational dynamics of syntaxin-3b from the ribbon synapses during assembly of the soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) complex.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages185-196
Number of pages12
DOIs
StatePublished - 2025

Publication series

NameMethods in Molecular Biology
Volume2887
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Munc18
  • Phosphorylation
  • SNARE proteins
  • Single-molecule FRET
  • Syntaxin-3b

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