Single-Molecule Manipulation Study of Chaperoned SNARE Folding and Assembly with Optical Tweezers

Huaizhou Jin, Jinghua Ge, Jie Yang, Yongli Zhang

Research output: Chapter in Book/Report/Conference proceedingChapter

1 Scopus citations

Abstract

Intracellular membrane fusion is primarily driven by coupled folding and assembly of soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs). SNARE assembly is intrinsically inefficient and must be chaperoned by a family of evolutionarily and structurally conserved Sec1/Munc-18 (SM) proteins. The physiological pathway of the chaperoned SNARE assembly has not been well understood, partly due to the difficulty in dissecting the many intermediates and pathways of SNARE assembly and measure their associated energetics and kinetics. Optical tweezers have proven to be a powerful tool to characterize the intermediates involved in the chaperoned SNARE assembly. Here, we demonstrate the application of optical tweezers combined with a homemade microfluidic system into studies of synaptic SNARE assembly chaperoned by their cognate SM protein Munc18-1. Three synaptic SNAREs and Munc18-1 constitute the core machinery for synaptic vesicle fusion involved in neurotransmitter release. Many other proteins further regulate the core machinery to enable fusion at the right time and location. The methods described here can be applied to other proteins that regulate SNARE assembly to control membrane fusion involved in numerous biological and physiological processes.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages461-481
Number of pages21
DOIs
StatePublished - 2022

Publication series

NameMethods in Molecular Biology
Volume2478
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics

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