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

Brazhe NA, Evlyukhin AB, Goodilin EA, Semenova AA, Novikov SM, Bozhevolnyi SI, Chichkov BN, Sarycheva AS, Baizhumanov AA, Nikelshparg EI, Deev LI, Maksimov EG, Maksimov GV, Sosnovtseva O - Sci Rep (2015)

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: Department of Biophysics, Biological Faculty, Moscow State University, Leninskie gory 1/12, Moscow, 119234, Russia.

ABSTRACT
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.

No MeSH data available.


Scheme of mitochondria placed on silver nanostructured surface (AgNSS).Schematic presentation of the magnified view of the outer mitochondrial membrane in a contact with AgNSS, intermembrane space (IMS) and internal mitochondrial membrane with complexes of the electron transport chain. Cytochromes of b and c-type are shown in corresponding complexes. Cytochrome c is shown as cyan ball. Red numbers indicate the approximate height of the outer mitochondrial membrane, size of IMS and IMS domain of complex III (data on distances and sizes of mitochondrial elements are taken from1129). VDAC–voltage-dependent anion channel; FCCP—protonophore (Carbonyl cyanide-p-trifluoromethoxyphenylhydrazone) used in the study.
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f1: Scheme of mitochondria placed on silver nanostructured surface (AgNSS).Schematic presentation of the magnified view of the outer mitochondrial membrane in a contact with AgNSS, intermembrane space (IMS) and internal mitochondrial membrane with complexes of the electron transport chain. Cytochromes of b and c-type are shown in corresponding complexes. Cytochrome c is shown as cyan ball. Red numbers indicate the approximate height of the outer mitochondrial membrane, size of IMS and IMS domain of complex III (data on distances and sizes of mitochondrial elements are taken from1129). VDAC–voltage-dependent anion channel; FCCP—protonophore (Carbonyl cyanide-p-trifluoromethoxyphenylhydrazone) used in the study.

Mentions: ETC of mitochondria includes cytochromes of b- (in complex II and III), c- (in complex III and cytochrome c) and a-types (in complex IV) (Fig. 1). Under used experimental conditions (532 nm laser), Raman scattering could be excited only from cytochromes of c- and b- types.


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

Brazhe NA, Evlyukhin AB, Goodilin EA, Semenova AA, Novikov SM, Bozhevolnyi SI, Chichkov BN, Sarycheva AS, Baizhumanov AA, Nikelshparg EI, Deev LI, Maksimov EG, Maksimov GV, Sosnovtseva O - Sci Rep (2015)

Scheme of mitochondria placed on silver nanostructured surface (AgNSS).Schematic presentation of the magnified view of the outer mitochondrial membrane in a contact with AgNSS, intermembrane space (IMS) and internal mitochondrial membrane with complexes of the electron transport chain. Cytochromes of b and c-type are shown in corresponding complexes. Cytochrome c is shown as cyan ball. Red numbers indicate the approximate height of the outer mitochondrial membrane, size of IMS and IMS domain of complex III (data on distances and sizes of mitochondrial elements are taken from1129). VDAC–voltage-dependent anion channel; FCCP—protonophore (Carbonyl cyanide-p-trifluoromethoxyphenylhydrazone) used in the study.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4561893&req=5

f1: Scheme of mitochondria placed on silver nanostructured surface (AgNSS).Schematic presentation of the magnified view of the outer mitochondrial membrane in a contact with AgNSS, intermembrane space (IMS) and internal mitochondrial membrane with complexes of the electron transport chain. Cytochromes of b and c-type are shown in corresponding complexes. Cytochrome c is shown as cyan ball. Red numbers indicate the approximate height of the outer mitochondrial membrane, size of IMS and IMS domain of complex III (data on distances and sizes of mitochondrial elements are taken from1129). VDAC–voltage-dependent anion channel; FCCP—protonophore (Carbonyl cyanide-p-trifluoromethoxyphenylhydrazone) used in the study.
Mentions: ETC of mitochondria includes cytochromes of b- (in complex II and III), c- (in complex III and cytochrome c) and a-types (in complex IV) (Fig. 1). Under used experimental conditions (532 nm laser), Raman scattering could be excited only from cytochromes of c- and b- types.

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: Department of Biophysics, Biological Faculty, Moscow State University, Leninskie gory 1/12, Moscow, 119234, Russia.

ABSTRACT
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.

No MeSH data available.