Quantifying interactions of a membrane protein embedded in a lipid nanodisc using fluorescence correlation spectroscopy

Sonny Ly, Feliza Bourguet, Nicholas O Fischer, Edmond Y Lau, Matthew A Coleman, Ted A. Laurence

Research output: Contribution to journalArticle

13 Scopus citations

Abstract

Using fluorescence correlation spectroscopy, we measured a dissociation constant of 20 nM between EGFP-labeled LcrV from Yersinia pestis and its cognate membrane-bound protein YopB inserted into a lipid nanodisc. The combination of fluorescence correlation spectroscopy and nanodisc technologies provides a powerful approach to accurately measure binding constants of interactions between membrane bound and soluble proteins in solution. Straightforward sample preparation, acquisition, and analysis procedures make this combined technology attractive for accurately measuring binding kinetics for this important class of protein-protein interactions.

Original languageEnglish (US)
JournalBiophysical Journal
Volume106
Issue number2
DOIs
StatePublished - Jan 21 2014

ASJC Scopus subject areas

  • Biophysics

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