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Optical spectroscopy in turbid media using an integrating sphere: mitochondrial chromophore analysis during metabolic transitions.
Anal Biochem 2013; 439(2):161-72AB

Abstract

Recent evidence suggests that the activity of mitochondrial oxidative phosphorylation complexes (MOPCs) is modulated at multiple sites. Here, a method of optically monitoring electron distribution within and between MOPCs is described using a center-mounted sample in an integrating sphere (to minimize scattering effects) with a rapid-scanning spectrometer. The redox-sensitive MOPC absorbances (∼465-630 nm) were modeled using linear least squares analysis with individual chromophore spectra. Classical mitochondrial activity transitions (e.g., ADP-induced increase in oxygen consumption) were used to characterize this approach. Most notable in these studies was the observation that intermediates of the catalytic cycle of cytochrome oxidase are dynamically modulated with metabolic state. The MOPC redox state, along with measurements of oxygen consumption and mitochondrial membrane potential, was used to evaluate the conductances of different sections of the electron transport chain. This analysis then was applied to mitochondria isolated from rabbit hearts subjected to ischemia/reperfusion (I/R). Surprisingly, I/R resulted in an inhibition of all measured MOPC conductances, suggesting a coordinated down-regulation of mitochondrial activity with this well-established cardiac perturbation.

Authors+Show Affiliations

Laboratory of Cardiac Energetics, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892, USA. chessdj@mail.nih.govNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, N.I.H., Intramural

Language

eng

PubMed ID

23665273

Citation

Chess, David J., et al. "Optical Spectroscopy in Turbid Media Using an Integrating Sphere: Mitochondrial Chromophore Analysis During Metabolic Transitions." Analytical Biochemistry, vol. 439, no. 2, 2013, pp. 161-72.
Chess DJ, Billings E, Covian R, et al. Optical spectroscopy in turbid media using an integrating sphere: mitochondrial chromophore analysis during metabolic transitions. Anal Biochem. 2013;439(2):161-72.
Chess, D. J., Billings, E., Covian, R., Glancy, B., French, S., Taylor, J., ... Balaban, R. S. (2013). Optical spectroscopy in turbid media using an integrating sphere: mitochondrial chromophore analysis during metabolic transitions. Analytical Biochemistry, 439(2), pp. 161-72. doi:10.1016/j.ab.2013.04.017.
Chess DJ, et al. Optical Spectroscopy in Turbid Media Using an Integrating Sphere: Mitochondrial Chromophore Analysis During Metabolic Transitions. Anal Biochem. 2013 Aug 15;439(2):161-72. PubMed PMID: 23665273.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Optical spectroscopy in turbid media using an integrating sphere: mitochondrial chromophore analysis during metabolic transitions. AU - Chess,David J, AU - Billings,Eric, AU - Covian,Raúl, AU - Glancy,Brian, AU - French,Stephanie, AU - Taylor,Joni, AU - de Bari,Heather, AU - Murphy,Elizabeth, AU - Balaban,Robert S, Y1 - 2013/05/09/ PY - 2013/02/25/received PY - 2013/04/03/revised PY - 2013/04/12/accepted PY - 2013/5/14/entrez PY - 2013/5/15/pubmed PY - 2013/11/12/medline KW - Electron transport chain KW - Ischemia/reperfusion KW - Mitochondria KW - Spectroscopy SP - 161 EP - 72 JF - Analytical biochemistry JO - Anal. Biochem. VL - 439 IS - 2 N2 - Recent evidence suggests that the activity of mitochondrial oxidative phosphorylation complexes (MOPCs) is modulated at multiple sites. Here, a method of optically monitoring electron distribution within and between MOPCs is described using a center-mounted sample in an integrating sphere (to minimize scattering effects) with a rapid-scanning spectrometer. The redox-sensitive MOPC absorbances (∼465-630 nm) were modeled using linear least squares analysis with individual chromophore spectra. Classical mitochondrial activity transitions (e.g., ADP-induced increase in oxygen consumption) were used to characterize this approach. Most notable in these studies was the observation that intermediates of the catalytic cycle of cytochrome oxidase are dynamically modulated with metabolic state. The MOPC redox state, along with measurements of oxygen consumption and mitochondrial membrane potential, was used to evaluate the conductances of different sections of the electron transport chain. This analysis then was applied to mitochondria isolated from rabbit hearts subjected to ischemia/reperfusion (I/R). Surprisingly, I/R resulted in an inhibition of all measured MOPC conductances, suggesting a coordinated down-regulation of mitochondrial activity with this well-established cardiac perturbation. SN - 1096-0309 UR - https://www.unboundmedicine.com/medline/citation/23665273/Optical_spectroscopy_in_turbid_media_using_an_integrating_sphere:_mitochondrial_chromophore_analysis_during_metabolic_transitions_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0003-2697(13)00191-7 DB - PRIME DP - Unbound Medicine ER -