Effect of heme iron valence state on the conformation of cytochrome c and its association with membrane interfaces. A CD and EPR investigation.
Recently cytochrome c has been mentioned as an important mediator in the events of cellular oxidative stress and apoptosis. To investigate the influence of charged interfaces on the conformation of cytochrome c, the CD and magnetic circular dichroic behavior of ferric and ferrous cytochrome c in homogeneous medium and in phosphatidylcholine/phosphatidylethanolamine/cardiolipin and dicetylphosphate liposomes was studied in the 300-600 and 200-320 nm wavelength region. EPR spectra demonstrate that the association of cytochrome c with membranes promotes alterations of the crystal field symmetry and spin state of the heme Fe(3+). The studies also include the effect of P(i), NaCl, and CaCl(2). Magnetic circular dichroism and CD results show that the interaction of both ferrous and ferric cytochrome c with charged interfaces promotes conformational changes in the alpha-helix content, tertiary structure, and heme iron spin state. Moreover, the association of cytochrome c with different liposomes is sensitive to the heme iron valence state. The more effective association with membranes occurs with ferrous cytochrome c. Dicetylphosphate liposomes, as a negatively charged membrane model, promoted a more pronounced conformational modification in the cytochrome c structure. A decrease in the lipid/protein association is detected in the presence of increasing amounts of CaCl(2), NaCl, and P(i), in response to the increase of the ionic strength.
Nantes
I L
IL
Centro Interdisciplinar de Investigação Bioquimica (CIIB), Prédio I, Sala 1S-15, Diretoria de Pesquisa e Pós-Graduação, Universidade de Mogi das Cruzes (UMC), CP 411, Mogi das Cruzes, São Paulo, CEP 08780-911, Brazil.
Zucchi
M R
MR
Nascimento
O R
OR
Faljoni-Alario
A
A
D000818:Animals; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism; D002942:Circular Dichroism; D003574:Cytochrome c Group / Q000737:chemistry* / Q000378:metabolism*; D003931:Diacetyl / Q000031:analogs & derivatives / Q000494:pharmacology; D004578:Electron Spin Resonance Spectroscopy; D006418:Heme / Q000737:chemistry / Q000378:metabolism*; D006736:Horses; D007501:Iron / Q000737:chemistry / Q000378:metabolism*; D008081:Liposomes / Q000737:chemistry / Q000378:metabolism*; D009206:Myocardium / Q000737:chemistry; D009943:Organophosphorus Compounds / Q000494:pharmacology; D009994:Osmolar Concentration; D017433:Protein Structure, Secondary; D012492:Salts / Q000494:pharmacology; D055672:Static Electricity
D003574:Cytochrome c Group; D008081:Liposomes; D009943:Organophosphorus Compounds; D012492:Salts; D006418:Heme; C005146:diacetyl phosphate; D007501:Iron; D003931:Diacetyl