Spin-label studies on the specificity of interaction of cardiolipin with beef heart cytochrome oxidase.
The selectivity of interaction of various cardiolipin analogues with beef heart cytochrome oxidase in reconstituted complexes with dimyristoylphosphatidylcholine has been studied by electron spin resonance spectroscopy, using lipids spin-labeled in the acyl chains. No difference in selectivity is observed between cardiolipin and its monolyso derivative, and similarly no selectivity is observed between phosphatidylcholine and lysophosphatidylcholine. Removal of the cardiolipin charge by methylation of the phosphate groups reduces but does not eliminate selectivity relative to phosphatidylcholine. The dependence of the lipid selectivity on head group and chain composition is in the order cardiolipin approximately equal to monolysocardiolipin greater than acylcardiolipin greater than dimethylcardiolipin greater than phosphatidylcholine approximately equal to lysophosphatidylcholine, where acylcardiolipin has the spin-label chain attached at the center -OH of the head group. The degree of association of the negatively charged cardiolipin derivatives with cytochrome oxidase decreases with increasing salt concentration, to a level comparable to that for dimethylcardiolipin. At high ionic strength there is still a marked selectivity relative to phosphatidylcholine. Li+ ions are more effective in screening the interaction than are Na+ ions, and divalent ions are more effective than monovalent ions. The selectivity for cardiolipin is only slightly reduced on titrating the protein to high pH. Alkylation of the protein with N-ethylmaleimide has little effect on the titration behavior. Covalent modification of the protein by reaction with citraconic anhydride decreases the selectivity of interaction with cardiolipin. It is concluded that cardiolipin possesses an additional specificity of interaction with cytochrome oxidase other than that of purely electrostatic origin.
Powell
G L
GL
Abteilung Spektroskopie, Max-Planck-Institut für biophysikalische Chemie, Göttingen, Federal Republic of Germany.
Knowles
P F
PF
Marsh
D
D
D000818:Animals; D002308:Cardiolipins / Q000378:metabolism*; D002417:Cattle; D004134:Dimyristoylphosphatidylcholine; D004578:Electron Spin Resonance Spectroscopy; D003576:Electron Transport Complex IV / Q000378:metabolism*; D007700:Kinetics; D009206:Myocardium / Q000201:enzymology*; D009994:Osmolar Concentration; D010713:Phosphatidylcholines / Q000378:metabolism; D013329:Structure-Activity Relationship
D002308:Cardiolipins; D010713:Phosphatidylcholines; D003576:Electron Transport Complex IV; D004134:Dimyristoylphosphatidylcholine