UNIT 19.12 Fluorescence Quenching Methods to Study Lipid-Protein Interactions
Published Online: 1 SEP 2006
Copyright © 2006 by John Wiley & Sons, Inc.
Lab Protocol Title
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.
- Published Online: 1 SEP 2006
- Published Print: AUG 2006
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.
- Lipid-protein interactions;
- membrane proteins;
- phospholipid synthesis;