Interaction of cytochrome c with mixed dimyristoylphosphatidylcholine-dimyristoylphosphatidylserine bilayers: a deuterium nuclear magnetic resonance study.
Deuterium nuclear magnetic resonance (2H NMR) was used to study the interaction of cytochrome c (from horse heart) with bilayers of mixed dimyristoylphosphatidylcholine (DMPC) and dimyristoylphosphatidylserine (DMPS). Three types of labeled lipids were used: chain-perdeuterated phosphatidylcholine (DMPC-d54), chain-perdeuterated phosphatidylserine (DMPS-d54), and phosphatidylserine labeled at the alpha-position of the head group (DMPS-d2). Liposomes containing equimolar mixtures of DMPC and DMPS were found to bind cytochrome c with a maximum ratio of about 1 mg of cytochrome c per 1 mg of DMPS. The 2H NMR spectra of equimolar mixtures of DMPC-d54-DMPS and DMPC-DMPS-d54 were examined with and without cytochrome c. No change of the NMR spectra of either DMPC or DMPS could be detected after protein addition, for temperatures both above and below the phospholipid phase transition region. On the other hand, in the liquid-crystalline state, the transverse relaxation time, T2e, was reduced by 30-40% after protein addition. Measurements of the spin-lattice relaxation time, T1, showed, under all circumstances, multiple components. For simplicity, we have examined the shape of the relaxation curves at short and long times. Addition of protein increased by 2-fold the value of the slow T1 component of DMPS-d54 but not that of DMPC-d54. Partially relaxed spectroscopy allowed us to assign this slow component (at least in part) to the methyl group and C2H2 groups near the methyl end of the chains, i.e., far from the binding sites of the extrinsic protein.(ABSTRACT TRUNCATED AT 250 WORDS)
D000818:Animals; D002152:Calorimetry, Differential Scanning / Q000379:methods; D003574:Cytochrome c Group / Q000378:metabolism*; D003903:Deuterium; D004134:Dimyristoylphosphatidylcholine / Q000378:metabolism*; D006736:Horses; D008051:Lipid Bilayers; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D008954:Models, Biological; D009206:Myocardium / Q000378:metabolism; D010718:Phosphatidylserines / Q000378:metabolism*; D011485:Protein Binding
D003574:Cytochrome c Group; D008051:Lipid Bilayers; D010718:Phosphatidylserines; C042859:dimyristoylphosphatidylserine; D003903:Deuterium; D004134:Dimyristoylphosphatidylcholine