Probing cytochrome c in living mitochondria with surface-enhanced Raman spectroscopy.
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.
Department of Biophysics, Biological Faculty, Moscow State University, Leninskie gory 1/12, Moscow, 119234, Russia.|Department of Biomedical Sciences, Faculty of Medical and Health Sciences, Copenhagen University, Blegdamsvej 3, Copenhagen, DK-2200, Denmark.
Nadezda A
NA
Brazhe
Laser Zentrum Hannover e. V., Hollerihallee 8, D-30419 Hannover, Germany.|Department of Technology and Innovation, University of Southern Denmark, Odense M, DK-5230 Denmark.
Andrey B
AB
Evlyukhin
Department of Nanomaterials, Faculty of Material Sciences, Moscow State University, Moscow, Leninskie gory 1/73, 119991, Russia.|Department of Inorganic chemistry, Faculty of Chemistry, Moscow State University, Moscow, Leninskie gory 1/3, 119991, Russia.|Kurnakov Institute of General and Inorganic Chemistry of Russian Academy of Sciences, Moscow, Leninskiy prospekt, 119992, Russia.
Eugene A
EA
Goodilin
Department of Nanomaterials, Faculty of Material Sciences, Moscow State University, Moscow, Leninskie gory 1/73, 119991, Russia.
Anna A
AA
Semenova
Department of Technology and Innovation, University of Southern Denmark, Odense M, DK-5230 Denmark.
Sergey M
SM
Novikov
Department of Technology and Innovation, University of Southern Denmark, Odense M, DK-5230 Denmark.
Sergey I
SI
Bozhevolnyi
Laser Zentrum Hannover e. V., Hollerihallee 8, D-30419 Hannover, Germany.
Boris N
BN
Chichkov
Department of Nanomaterials, Faculty of Material Sciences, Moscow State University, Moscow, Leninskie gory 1/73, 119991, Russia.
Asya S
AS
Sarycheva
Department of Biophysics, Biological Faculty, Moscow State University, Leninskie gory 1/12, Moscow, 119234, Russia.
Adil A
AA
Baizhumanov
Department of Biophysics, Biological Faculty, Moscow State University, Leninskie gory 1/12, Moscow, 119234, Russia.
Evelina I
EI
Nikelshparg
Department of Biophysics, Biological Faculty, Moscow State University, Leninskie gory 1/12, Moscow, 119234, Russia.
Leonid I
LI
Deev
Department of Biophysics, Biological Faculty, Moscow State University, Leninskie gory 1/12, Moscow, 119234, Russia.
Eugene G
EG
Maksimov
Department of Biophysics, Biological Faculty, Moscow State University, Leninskie gory 1/12, Moscow, 119234, Russia.
Georgy V
GV
Maksimov
Department of Biomedical Sciences, Faculty of Medical and Health Sciences, Copenhagen University, Blegdamsvej 3, Copenhagen, DK-2200, Denmark.
Olga
O
Sosnovtseva
D000255:Adenosine Triphosphate / Q000096:biosynthesis; D000818:Animals; D045304:Cytochromes c / Q000737:chemistry* / Q000378:metabolism*; D004579:Electron Transport; D008297:Male; D053078:Membrane Potential, Mitochondrial; D008928:Mitochondria / Q000378:metabolism*; D008929:Mitochondria, Heart / Q000378:metabolism; D011522:Protons; D051381:Rats; D013059:Spectrum Analysis, Raman / Q000379:methods
D011522:Protons; D000255:Adenosine Triphosphate; D045304:Cytochromes c