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pH-Induced Switch between Different Modes of Cytochrome c Binding to Cardiolipin-Containing Liposomes
Journal article   Open access   Peer reviewed

pH-Induced Switch between Different Modes of Cytochrome c Binding to Cardiolipin-Containing Liposomes

Bridget Milorey, Reinhard Schweitzer-Stenner, Raghed Kurbaj and Dmitry Malyshka
ACS omega, v 4(1), pp 1386-1400
01 Jan 2019
PMID: 31459406
url
https://doi.org/10.1021/acsomega.8b02574View
Published, Version of Record (VoR)Open Access (Publisher-Specific) Open

Abstract

Chemistry Chemistry, Multidisciplinary Physical Sciences Science & Technology
Fluorescence, visible circular dichroism (CD), absorption, and resonance Raman spectroscopy techniques were combined to explore structural changes of ferricytochrome c upon its binding to cardiolipin-containing liposomes (20% 1,1', 1,2'-tetraoleyolcardiolipin and 1,2-deoleyol-sn-glycero-3-phosphocholine) at acidic pH (6.5). According to the earlier work of Kawai et al. [J. Biol. Chem. 2005, 280, 34709-347171], cytochrome c binding at this pH is governed by interactions between the phosphate head groups of cardiolipin and amino acid side chains of the so-called L-site, which contains the charged residues K22, K25, K27, and potentially H26 and H33. We found that L-site binding causes a conformational transition that involves a change of the protein's ligation and spin state. In this paper, we report spectroscopic responses to an increasing number of cardiolipin-containing liposomes at pH 6.5 in the absence and presence of NaCl. The latter was found to mostly inhibit protein binding already with 50 mM concentration. The inhibition effect can be quantitatively reproduced by applying the electrostatic theory of Heimburg et al. [Biophys. J. 1995, 68, 536-546]. A comparison with corresponding spectroscopic response data obtained at pH 7.4 reveals major differences in that the latter indicates hydrophobic binding, followed by an electrostatically driven conformational change. Visible CD data suggest that structural changes in the heme pocket of liposome-bound ferricytochrome c resemble to some extent those in the denatured protein in urea at neutral and acidic pH. The measured noncoincidence between absorption and CD Soret band of cytochrome c in the presence of a large access of cardiolipin is caused by the electric field at the membrane surface. The very fact that its contribution to the internal electric field in the heme pocket is detectable by spectroscopic means suggests some penetration of the protein into membrane surface.

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Web of Science research areas
Chemistry, Multidisciplinary
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