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Probing cytochrome c in living mitochondria with surface-enhanced Raman spectroscopy

Nadezda A. BrazheDepartment of Biomedical Sciences, Faculty of Medical and Health Sciences, Copenhagen University, Blegdamsvej 3, Copenhagen, DK-2200, DenmarkAndrey B. EvlyukhinDepartment of Technology and Innovation, University of Southern Denmark, Odense M, DK-5230 DenmarkEugene A. GoodilinDepartment of Inorganic chemistry, Faculty of Chemistry, Moscow State University, Moscow, Leninskie gory 1/3, 119991, RussiaAnna A. SemenovaDepartment of Nanomaterials, Faculty of Material Sciences, Moscow State University, Moscow, Leninskie gory 1/73, 119991, RussiaSergey M. NovikovDepartment of Technology and Innovation, University of Southern Denmark, Odense M, DK-5230 DenmarkSergey I. BozhevolnyiDepartment of Technology and Innovation, University of Southern Denmark, Odense M, DK-5230 DenmarkBoris N. ChichkovLaser Zentrum Hannover e. V., Hollerihallee 8, D-30419 Hannover, GermanyAsya S. SarychevaDepartment of Nanomaterials, Faculty of Material Sciences, Moscow State University, Moscow, Leninskie gory 1/73, 119991, RussiaА. А. БайжумановDepartment of Biophysics, Biological Faculty, Moscow State University, Leninskie gory 1/12, Moscow, 119234, RussiaEvelina I. NikelshpargDepartment of Biophysics, Biological Faculty, Moscow State University, Leninskie gory 1/12, Moscow, 119234, RussiaЛ. И. ДеевDepartment of Biophysics, Biological Faculty, Moscow State University, Leninskie gory 1/12, Moscow, 119234, RussiaEugene G. MaksimovDepartment of Biophysics, Biological Faculty, Moscow State University, Leninskie gory 1/12, Moscow, 119234, RussiaГ. В. МаксимовDepartment of Biophysics, Biological Faculty, Moscow State University, Leninskie gory 1/12, Moscow, 119234, RussiaOlga SosnovtsevaDepartment of Biomedical Sciences, Faculty of Medical and Health Sciences, Copenhagen University, Blegdamsvej 3, Copenhagen, DK-2200, Denmark

2015en

ABI

Annotatsiya

Selective study of the electron transport chain components in living mitochondria is essential for fundamental biophysical research and for the development of new medical diagnostic methods. However, many important details of inter- and intramembrane mitochondrial processes have remained in shadow due to the lack of non-invasive techniques. Here we suggest a novel label-free approach based on the surface-enhanced Raman spectroscopy (SERS) to monitor the redox state and conformation of cytochrome c in the electron transport chain in living mitochondria. We demonstrate that SERS spectra of living mitochondria placed on hierarchically structured silver-ring substrates provide exclusive information about cytochrome c behavior under modulation of inner mitochondrial membrane potential, proton gradient and the activity of ATP-synthetase. Mathematical simulation explains the observed enhancement of Raman scattering due to high concentration of electric near-field and large contact area between mitochondria and nanostructured surfaces.

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DOI: 10.1038/srep13793

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