Unit

UNIT 19.12 Fluorescence Quenching Methods to Study Lipid-Protein Interactions

  1. Joanne Carney,
  2. J. Malcolm East,
  3. Sanjay Mall,
  4. Phedra Marius,
  5. Andrew M. Powl,
  6. J. Neville Wright,
  7. Anthony G. Lee

Published Online: 1 SEP 2006

DOI: 10.1002/0471142301.ps1912s45

Current Protocols in Protein Science

Current Protocols in Protein Science

How to Cite

Carney, J., East, J. M., Mall, S., Marius, P., Powl, A. M., Wright, J. N. and Lee, A. G. 2006. Fluorescence Quenching Methods to Study Lipid-Protein Interactions. Current Protocols in Protein Science. 45:19.12:19.12.1–19.12.17.

Author Information

  1. University of Southampton, Southampton, United Kingdom

Publication History

  1. Published Online: 1 SEP 2006
  2. Published Print: AUG 2006

Abstract

This unit describes how fluorescence quenching methods can be used to determine binding constants for phospholipids binding to intrinsic membrane proteins. Reconstitution of a Trp-containing intrinsic membrane protein with bromine-containing phospholipids leads to quenching of the Trp fluorescence of the protein; the extent of quenching depends on the strength of binding of the phospholipid to the protein. Protocols are included for the synthesis of bromine-containing phospholipids from phospholipids containing carbon-carbon double bonds in their fatty acyl chains and for the reconstitution of membrane proteins into bilayers containing bromine-containing phospholipids. Details are included on data analysis, including equations and software that can be used for fitting the fluorescence quenching data.

Keywords:

  • Lipid-protein interactions;
  • reconstitution;
  • membrane proteins;
  • fluorescence;
  • phospholipid synthesis;
  • bromination