Fluorescence Assays to Study Membrane Penetration of Proteins

Yosuke Senju, Hongxia Zhao

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Phosphoinositides play important roles in the regulation of protein recruitment at specialized membrane domains, protein activity, and membrane dynamics. Phosphoinositide–protein interplay occurs via multiple mechanisms and proteins associate with membranes through different binding patterns. Determinations of membrane-binding mode and membrane penetration depth of proteins in lipid bilayer are thus important steps in characterizing the molecular mechanisms of membrane–protein interactions. Here, we show two standard in vitro assays using liposomes, diphenylhexatriene (DPH) anisotropy, and fluorescence quenching by brominated lipids to determine membrane penetration of proteins into lipid bilayer. These methods will provide useful tools to study membrane–protein association and uncover molecular details of protein–lipid interplay, which are important for understanding biological functions of membrane-associated proteins and membrane dynamics.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages215-223
Number of pages9
DOIs
Publication statusPublished - 2021

Publication series

NameMethods in Molecular Biology
Volume2251
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Anisotropy
  • Lipid–protein interactions
  • Liposome
  • Membrane insertion
  • Membrane penetration
  • Phosphoinositides

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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