title: Multiple binding sites for fatty acids on the potassium channel KcsA.
abstract: Interactions of fatty acids with the potassium channel KcsA were studied using Trp fluorescence quenching and electron paramagnetic resonance (EPR) techniques. The brominated analogue of oleic acid was shown to bind to annular sites on KcsA and to the nonannular sites at each protein-protein interface in the homotetrameric structure with binding constants relative to dioleoylphosphatidylcholine of 0.67 ± 0.04 and 0.87 ± 0.08, respectively. Mutation of the two Arg residues close to the nonannular binding sites had no effect on fatty acid binding. EPR studies with a spin-labeled analogue of stearic acid detected a high-affinity binding site for the fatty acid with strong immobilization. Fluorescence quenching studies with the spin-labeled analogue showed that the binding site detected in the EPR experiments could not be one of the annular or nonannular binding sites. Instead, it is proposed that the EPR studies detect binding to the central hydrophobic cavity of the channel, with a binding constant in the range of ~0.1-1 μM.
authors.affiliation: Centre for Biological Sciences, Life Sciences Building, University of Southampton, Southampton SO17 1BJ, UK.
authors.forename: Juan H
authors.initials: JH
authors.lastname: Bolivar
authors.forename: Natalie
authors.initials: N
authors.lastname: Smithers
authors.forename: J Malcolm
authors.initials: JM
authors.lastname: East
authors.forename: Derek
authors.initials: D
authors.lastname: Marsh
authors.forename: Anthony G
authors.initials: AG
authors.lastname: Lee
mesh_terms: D001665:Binding Sites; D004578:Electron Spin Resonance Spectroscopy; D005227:Fatty Acids / Q000378:metabolism*; D015221:Potassium Channels / Q000378:metabolism*
chemical_list: D005227:Fatty Acids; D015221:Potassium Channels