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SNAREs pp 237-249 | Cite as

Functional Reconstitution of Intracellular Vesicle Fusion Using Purified SNAREs and Sec1/Munc18 (SM) Proteins

  • Haijia Yu
  • Lauren Crisman
  • Michael H. B. Stowell
  • Jingshi Shen
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1860)

Abstract

The fusion of intracellular vesicles with target membranes is mediated by two classes of conserved molecules—soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNAP receptors or SNAREs) and Sec1/Munc18 (SM) proteins. A conserved function of SM proteins is to recognize their cognate trans-SNARE complexes and accelerate fusion kinetics. Here, we describe a physiologically relevant reconstitution system in which macromolecular crowding agents are included to recapitulate the crowded intracellular environment. Through this system, we elucidate the molecular mechanisms by which SNAREs and SM proteins drive vesicle fusion.

Key words

SNARE SM protein Vesicle fusion Membrane fusion Reconstitution Lipid mixing Content mixing Macromolecular crowding 

Notes

Acknowledgments

This work was supported by National Institutes of Health grants GM102217 and DK095367 (JS).

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Jiangsu Key Laboratory for Molecular and Medical Biotechnology, College of Life SciencesNanjing Normal UniversityNanjingChina
  2. 2.Department of Molecular, Cellular and Developmental BiologyUniversity of ColoradoBoulderUSA

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