The interaction between cytochrome c (Cyt c) and cardiolipin (CL) plays a vital role in the early stages of apoptosis. The binding of CL to Cyt c induces a considerable increase in its peroxidase activity that has been attributed to the partial unfolding of the protein, dissociation of the Met80 axial ligand, and formation of non-native conformers. Although the interaction between Cyt c and CL has been extensively studied, there is still no consensus regarding the conformational rearrangements of Cyt c that follow the protein-lipid interaction. To rationalize the different results and gain better insight into the Cyt c-CL interaction, we have studied the formation of the CL complex of the horse heart wild-type protein and selected mutants in which residues considered to play a key role in the interaction with CL (His26, His33, Lys72, Lys73, and Lys79) have been mutated. The analysis was conducted at both room temperature and low temperatures via ultraviolet-visible absorption, resonance Raman, and electron paramagnetic resonance spectroscopies. The trigger and the sequence of CL-induced structural variations are discussed in terms of disruption of the His26-Pro44 hydrogen bond. We unequivocally identify the sixth ligand in the partially unfolded, non-native low-spin state that Cyt c can adopt following the protein-lipid interaction, as a His ligation, ruling out the previously proposed involvement of a Lys residue or an OH- ion.
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|Titolo:||Unravelling the non-native low-spin state of the cytochrome c-cardiolipin complex: evidence of the formation of a His-ligated species only|
|Citazione:||Milazzo, L., Tognaccini, L., Howes, B.D., Sinibaldi, F., Piro, M.C., Fittipaldi, M., et al. (2017). Unravelling the non-native low-spin state of the cytochrome c-cardiolipin complex: evidence of the formation of a His-ligated species only. BIOCHEMISTRY, 56(13), 1887-1898.|
|Appare nelle tipologie:||1.1 Articolo in rivista|
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