Nonlinear electron paramagnetic resonance studies of the interaction of cytochrome c oxidase with spin-labeled lipids in gel-phase membranes.
The interaction of lipids, spin-labeled at different positions in the sn-2 chain, with cytochrome c oxidase reconstituted in gel-phase membranes of dimyristoylphosphatidylglycerol has been studied by electron paramagnetic resonance (EPR) spectroscopy. Nonlinear EPR methods, both saturation transfer EPR and progressive saturation EPR, were used. Interaction with the protein largely removes the flexibility gradient of the lipid chains in gel-phase membranes. The rotational mobility of the chain segments is reduced, relative to that for gel-phase lipids, by the intramembranous interaction with cytochrome c oxidase. This holds for all positions of chain labeling, but the relative effect is greater for chain segments closer to the terminal methyl ends. Modification of the paramagnetic metal-ion centers in the protein by binding azide has a pronounced effect on the spin-lattice relaxation of the lipid spin labels. This demonstrates that the centers modified are sufficiently close to the first-shell lipids to give appreciable dipolar interactions and that their vertical location in the membrane is closer to the 5-position than to the 14-position of the lipid chains.
Páli
T
T
Max-Planck-Institut für biophysikalische Chemie, Abteilung Spektroskopie, D-37070 Göttingen, Germany.
Kleinschmidt
J H
JH
Powell
G L
GL
Marsh
D
D
D000818:Animals; D016880:Anisotropy; D001386:Azides / Q000737:chemistry; D001665:Binding Sites; D002417:Cattle; D003300:Copper / Q000737:chemistry; D004578:Electron Spin Resonance Spectroscopy / Q000379:methods; D003576:Electron Transport Complex IV / Q000737:chemistry*; D006418:Heme / Q000737:chemistry; D008563:Membrane Lipids / Q000737:chemistry*; D010715:Phosphatidylglycerols / Q000737:chemistry*; D013113:Spin Labels
D001386:Azides; D008563:Membrane Lipids; D010715:Phosphatidylglycerols; D013113:Spin Labels; D006418:Heme; D003300:Copper; C002773:dimyristoylphosphatidylglycerol; D003576:Electron Transport Complex IV