title: Fluorescence quenching methods to study lipid-protein interactions.
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.
authors.affiliation: University of Southampton, Southampton, United Kingdom.
authors.forename: Joanne
authors.initials: J
authors.lastname: Carney
authors.forename: J Malcolm
authors.initials: JM
authors.lastname: East
authors.forename: Sanjay
authors.initials: S
authors.lastname: Mall
authors.forename: Phedra
authors.initials: P
authors.lastname: Marius
authors.forename: Andrew M
authors.initials: AM
authors.lastname: Powl
authors.forename: J Neville
authors.initials: JN
authors.lastname: Wright
authors.forename: Anthony G
authors.initials: AG
authors.lastname: Lee
mesh_terms: D001966:Bromine / Q000737:chemistry; D005453:Fluorescence; D013058:Mass Spectrometry; D008565:Membrane Proteins / Q000737:chemistry* / Q000378:metabolism; D010743:Phospholipids / Q000737:chemistry* / Q000378:metabolism; D011485:Protein Binding
chemical_list: D008565:Membrane Proteins; D010743:Phospholipids; D001966:Bromine