Single-vesicle measurement of protein-induced membrane tethering

Bin Cai, Luning Yu, Savanna R. Sharum, Kai Zhang, Jiajie Diao

Research output: Contribution to journalArticlepeer-review

Abstract

Functions of the proteins involved in membrane tethering, a crucial step in membrane trafficking, remain elusive due to the lack of effective tools to investigate protein-lipid interaction. To address this challenge, we introduce a method to study protein-induced membrane tethering via in vitro reconstitution of lipid vesicles, including detailed steps from the preparation of the PEGylated slides to the imaging of single vesicles. Furthermore, we demonstrate the measurement of protein-vesicle interaction in tethered vesicle pairs using two representative proteins, the cytoplasmic domain of synaptotagmin-1 (C2AB) and α-synuclein. Results from Förster (fluorescence) resonance energy transfer (FRET) reveal that membrane tethering is distinguished from membrane fusion. Single-vesicle measurement also allows for assessment of dose-dependent effects of proteins and ions on membrane tethering. We envision that the continuous development of advanced techniques in the single-vesicle measurement will enable the investigation of complex protein-membrane interactions in live cells or tissues.

Original languageEnglish (US)
Pages (from-to)267-273
Number of pages7
JournalColloids and Surfaces B: Biointerfaces
Volume177
DOIs
StatePublished - May 1 2019

ASJC Scopus subject areas

  • Biotechnology
  • Surfaces and Interfaces
  • Physical and Theoretical Chemistry
  • Colloid and Surface Chemistry

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