Monitoring the location profile of fluorophores in phosphatidylcholine bilayers by the use or paramagnetic quenching.
Spin probes differing in the position of their paramagnetic centre are used to quench the fluorescence of pyrene derivatives and chlorophylls incorporated into dimyristoyl phosphatidylcholine membranes. Pyrene butyric acid and pyrene decanoic acid with known orientation relative to the membrane surface are investigated. The quenching efficiency of fatty acid spin probes is dependent on the position of the nitroxide radical group in the fatty acid chain. Using this short range interaction we developed a spectroscopic method to chlorophyll-containing vesicles, we were able to characterize the orientation of the porphyrin ring within the membrane. Moreover, the chlorophyll fluorescence is also quenched by a water-soluble spin label. Therefore the porphyrin ring appears to be orientated in the polar head group region of the lipid layer, but not to be protruding out into the water phase. This conclusion is confirmed by the use of pyrene derivatives. Fluorescence quenching by a water-soluble spin label within the lipid matrix is observed even in the rigid state of the membrane. Fluorescence lifetime measurements suggest the existence of two different quenching mechanisms: (1) a static quenching occurring below the lipid phase transition temperature, and (2) an additional dynamic quenching taking place in the fluid state of the lipid bilayer.
D002734:Chlorophyll; D004578:Electron Spin Resonance Spectroscopy; D008567:Membranes, Artificial; D008968:Molecular Conformation; D010713:Phosphatidylcholines; D011721:Pyrenes; D013050:Spectrometry, Fluorescence; D013113:Spin Labels
D008567:Membranes, Artificial; D010713:Phosphatidylcholines; D011721:Pyrenes; D013113:Spin Labels; D002734:Chlorophyll