Active Site Structure and Peroxidase Activity of Oxidatively Modified Cytochrome c Species in Complexes with Cardiolipin

We report a resonance Raman and UV-vis characterization of the active site structure of oxidatively modified forms of cytochrome c (Cyt-c) free in solution and in complexes with cardiolipin (CL). The studied post-translational modifications of Cyt-c include methionine sulfoxidation and tyrosine nitr...

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Autores principales:	Capdevila, D.A., Oviedo Rouco, S., Tomasina, F., Tortora, V., Demicheli, V., Radi, R., Murgida, D.H.
Formato:	JOUR
Materias:	Amino acids Binding energy Bins Enzyme activity Liposomes Nitration Oxidation Phospholipids Active site structure Enzymatic activities Lipid peroxidation Peroxidase activities Post-translational modifications Proapoptotic signals Tyrosine nitration Wild-type proteins Proteins cardiolipin cytochrome c heme liposome methionine peroxidase protein variant tyrosine Article conformational transition controlled study enzyme active site enzyme activity enzyme structure excitation lipid peroxidation nitration oxidative stress priority journal protein processing protein protein interaction Raman spectrometry sulfoxidation ultracentrifugation ultraviolet spectroscopy animal chemistry horse protein conformation ultraviolet spectrophotometry Animals Cardiolipins Catalytic Domain Cytochromes c Horses Protein Conformation Spectrophotometry, Ultraviolet Spectrum Analysis, Raman
Acceso en línea:	http://hdl.handle.net/20.500.12110/paper_00062960_v54_n51_p7491_Capdevila
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

id	todo:paper_00062960_v54_n51_p7491_Capdevila
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spelling	todo:paper_00062960_v54_n51_p7491_Capdevila2023-10-03T14:04:36Z Active Site Structure and Peroxidase Activity of Oxidatively Modified Cytochrome c Species in Complexes with Cardiolipin Capdevila, D.A. Oviedo Rouco, S. Tomasina, F. Tortora, V. Demicheli, V. Radi, R. Murgida, D.H. Amino acids Binding energy Bins Enzyme activity Liposomes Nitration Oxidation Phospholipids Active site structure Enzymatic activities Lipid peroxidation Peroxidase activities Post-translational modifications Proapoptotic signals Tyrosine nitration Wild-type proteins Proteins cardiolipin cytochrome c heme liposome methionine peroxidase protein variant tyrosine cardiolipin cytochrome c Article conformational transition controlled study enzyme active site enzyme activity enzyme structure excitation lipid peroxidation nitration oxidative stress priority journal protein processing protein protein interaction Raman spectrometry sulfoxidation ultracentrifugation ultraviolet spectroscopy animal chemistry enzyme active site horse protein conformation ultraviolet spectrophotometry Animals Cardiolipins Catalytic Domain Cytochromes c Horses Protein Conformation Spectrophotometry, Ultraviolet Spectrum Analysis, Raman We report a resonance Raman and UV-vis characterization of the active site structure of oxidatively modified forms of cytochrome c (Cyt-c) free in solution and in complexes with cardiolipin (CL). The studied post-translational modifications of Cyt-c include methionine sulfoxidation and tyrosine nitration, which lead to altered heme axial ligation and increased peroxidase activity with respect to those of the wild-type protein. In spite of the structural and activity differences between the protein variants free in solution, binding to CL liposomes induces in all cases the formation of a spectroscopically identical bis-His axial coordination conformer that more efficiently promotes lipid peroxidation. The spectroscopic results indicate that the bis-His form is in equilibrium with small amounts of high-spin species, thus suggesting a labile distal His ligand as the basis for the CL-induced increase in enzymatic activity observed for all protein variants. For Cyt-c nitrated at Tyr74 and sulfoxidized at Met80, the measured apparent binding affinities for CL are ∼4 times larger than for wild-type Cyt-c. On the basis of these results, we propose that these post-translational modifications may amplify the pro-apoptotic signal of Cyt-c under oxidative stress conditions at CL concentrations lower than for the unmodified protein. © 2015 American Chemical Society. Fil:Capdevila, D.A. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Murgida, D.H. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_00062960_v54_n51_p7491_Capdevila
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-134
collection	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic	Amino acids Binding energy Bins Enzyme activity Liposomes Nitration Oxidation Phospholipids Active site structure Enzymatic activities Lipid peroxidation Peroxidase activities Post-translational modifications Proapoptotic signals Tyrosine nitration Wild-type proteins Proteins cardiolipin cytochrome c heme liposome methionine peroxidase protein variant tyrosine cardiolipin cytochrome c Article conformational transition controlled study enzyme active site enzyme activity enzyme structure excitation lipid peroxidation nitration oxidative stress priority journal protein processing protein protein interaction Raman spectrometry sulfoxidation ultracentrifugation ultraviolet spectroscopy animal chemistry enzyme active site horse protein conformation ultraviolet spectrophotometry Animals Cardiolipins Catalytic Domain Cytochromes c Horses Protein Conformation Spectrophotometry, Ultraviolet Spectrum Analysis, Raman
spellingShingle	Amino acids Binding energy Bins Enzyme activity Liposomes Nitration Oxidation Phospholipids Active site structure Enzymatic activities Lipid peroxidation Peroxidase activities Post-translational modifications Proapoptotic signals Tyrosine nitration Wild-type proteins Proteins cardiolipin cytochrome c heme liposome methionine peroxidase protein variant tyrosine cardiolipin cytochrome c Article conformational transition controlled study enzyme active site enzyme activity enzyme structure excitation lipid peroxidation nitration oxidative stress priority journal protein processing protein protein interaction Raman spectrometry sulfoxidation ultracentrifugation ultraviolet spectroscopy animal chemistry enzyme active site horse protein conformation ultraviolet spectrophotometry Animals Cardiolipins Catalytic Domain Cytochromes c Horses Protein Conformation Spectrophotometry, Ultraviolet Spectrum Analysis, Raman Capdevila, D.A. Oviedo Rouco, S. Tomasina, F. Tortora, V. Demicheli, V. Radi, R. Murgida, D.H. Active Site Structure and Peroxidase Activity of Oxidatively Modified Cytochrome c Species in Complexes with Cardiolipin
topic_facet	Amino acids Binding energy Bins Enzyme activity Liposomes Nitration Oxidation Phospholipids Active site structure Enzymatic activities Lipid peroxidation Peroxidase activities Post-translational modifications Proapoptotic signals Tyrosine nitration Wild-type proteins Proteins cardiolipin cytochrome c heme liposome methionine peroxidase protein variant tyrosine cardiolipin cytochrome c Article conformational transition controlled study enzyme active site enzyme activity enzyme structure excitation lipid peroxidation nitration oxidative stress priority journal protein processing protein protein interaction Raman spectrometry sulfoxidation ultracentrifugation ultraviolet spectroscopy animal chemistry enzyme active site horse protein conformation ultraviolet spectrophotometry Animals Cardiolipins Catalytic Domain Cytochromes c Horses Protein Conformation Spectrophotometry, Ultraviolet Spectrum Analysis, Raman
description	We report a resonance Raman and UV-vis characterization of the active site structure of oxidatively modified forms of cytochrome c (Cyt-c) free in solution and in complexes with cardiolipin (CL). The studied post-translational modifications of Cyt-c include methionine sulfoxidation and tyrosine nitration, which lead to altered heme axial ligation and increased peroxidase activity with respect to those of the wild-type protein. In spite of the structural and activity differences between the protein variants free in solution, binding to CL liposomes induces in all cases the formation of a spectroscopically identical bis-His axial coordination conformer that more efficiently promotes lipid peroxidation. The spectroscopic results indicate that the bis-His form is in equilibrium with small amounts of high-spin species, thus suggesting a labile distal His ligand as the basis for the CL-induced increase in enzymatic activity observed for all protein variants. For Cyt-c nitrated at Tyr74 and sulfoxidized at Met80, the measured apparent binding affinities for CL are ∼4 times larger than for wild-type Cyt-c. On the basis of these results, we propose that these post-translational modifications may amplify the pro-apoptotic signal of Cyt-c under oxidative stress conditions at CL concentrations lower than for the unmodified protein. © 2015 American Chemical Society.
format	JOUR
author	Capdevila, D.A. Oviedo Rouco, S. Tomasina, F. Tortora, V. Demicheli, V. Radi, R. Murgida, D.H.
author_facet	Capdevila, D.A. Oviedo Rouco, S. Tomasina, F. Tortora, V. Demicheli, V. Radi, R. Murgida, D.H.
author_sort	Capdevila, D.A.
title	Active Site Structure and Peroxidase Activity of Oxidatively Modified Cytochrome c Species in Complexes with Cardiolipin
title_short	Active Site Structure and Peroxidase Activity of Oxidatively Modified Cytochrome c Species in Complexes with Cardiolipin
title_full	Active Site Structure and Peroxidase Activity of Oxidatively Modified Cytochrome c Species in Complexes with Cardiolipin
title_fullStr	Active Site Structure and Peroxidase Activity of Oxidatively Modified Cytochrome c Species in Complexes with Cardiolipin
title_full_unstemmed	Active Site Structure and Peroxidase Activity of Oxidatively Modified Cytochrome c Species in Complexes with Cardiolipin
title_sort	active site structure and peroxidase activity of oxidatively modified cytochrome c species in complexes with cardiolipin
url	http://hdl.handle.net/20.500.12110/paper_00062960_v54_n51_p7491_Capdevila
work_keys_str_mv	AT capdevilada activesitestructureandperoxidaseactivityofoxidativelymodifiedcytochromecspeciesincomplexeswithcardiolipin AT oviedoroucos activesitestructureandperoxidaseactivityofoxidativelymodifiedcytochromecspeciesincomplexeswithcardiolipin AT tomasinaf activesitestructureandperoxidaseactivityofoxidativelymodifiedcytochromecspeciesincomplexeswithcardiolipin AT tortorav activesitestructureandperoxidaseactivityofoxidativelymodifiedcytochromecspeciesincomplexeswithcardiolipin AT demicheliv activesitestructureandperoxidaseactivityofoxidativelymodifiedcytochromecspeciesincomplexeswithcardiolipin AT radir activesitestructureandperoxidaseactivityofoxidativelymodifiedcytochromecspeciesincomplexeswithcardiolipin AT murgidadh activesitestructureandperoxidaseactivityofoxidativelymodifiedcytochromecspeciesincomplexeswithcardiolipin
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Active Site Structure and Peroxidase Activity of Oxidatively Modified Cytochrome c Species in Complexes with Cardiolipin

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