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The determination of the composition of the complex of cytochrome c with cardiolipin using spectrophotometric titration and thermal lens spectrometry


E.V. Proskurnina – Ph.D. (Chem.), Associate Professor, Department of Medical Biophysics, Faculty of Fundamental Medicine, Lomonosov Moscow State University
M.A. Proskurnin – D.Sc.(Chem.), Professor, Department of Analytical Chemistry, Chemical Faculty, Lomonosov Moscow State University
A.V. Alekseev – Ph.D. (Biol.), Senior Research Scientist, Russian Research Institute of Aviation Materials
V.R. Galimova – Student, Department of Analytical Chemistry, Chemical Faculty, Lomonosov Moscow State University. Moscow
Yu.A. Vladimirov – Dr. Sc. (Biol.), Professor, Full Member of the Russian Academy of Sciences, Head of the Department of Medical Biophysics, Faculty of Fundamental Medicine, Lomonosov Moscow State University, Lomonosovsky

Using spectrophotometric titration and thermal-lens spectrometry, the composition of cytochrome c-cardiolipin complex was determined. This complex plays a key role in mitochondrial way of apoptosis. Spectrophotometric titration was performed in the range of micromolar concentrations, where complex of cytochrome c and cardiolipin precipitated. Thermal-lens measurements were performed at nanomolar concentrations, where the complex is soluble. Cytochrome c : cardiolipin ratio was determined as 59±7 at pH 7,4; 38±5 at pH 6,5, and 13±3 at pH 3,7 from the spectrophotometric data. Cytochrome c : cardiolipin ratio was determined as 60 at pH 7,4; 30 at pH 6,8, and 10 at pH 5,5 from the thermal-lens data. For pH 7.4 the constant of molecular solubility was calculated, 30 μmol/L.

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