For Publishers
DiscoveryMetadataPeer reviewHostingPublishing
For Researchers
JoinPublishReviewCollect
Register
Blog
About
Search
Advanced search
My ScienceOpen
Search
For Publishers
For Researchers
Blog
About
41
views
28
references
 Top references
cited by
222
    
0 reviews
 Review
0
comments
 Comment
0
recommends
+1 Recommend
0 collections
    0
    shares
      0
      similar
       All similar
      • Record: found
      • Abstract: found
      • Article: found
      Is Open Access

      Sites of reactive oxygen species generation by mitochondria oxidizing different substrates ☆

      research-article

      Read this article at

      ScienceOpenPublisherPMC
      Bookmark
          There is no author summary for this article yet. Authors can add summaries to their articles on ScienceOpen to make them more accessible to a non-specialist audience.

          Abstract

          Mitochondrial radical production is important in redox signaling, aging and disease, but the relative contributions of different production sites are poorly understood. We analyzed the rates of superoxide/H 2O 2 production from different defined sites in rat skeletal muscle mitochondria oxidizing a variety of conventional substrates in the absence of added inhibitors: succinate; glycerol 3-phosphate; palmitoylcarnitine plus carnitine; or glutamate plus malate. In all cases, the sum of the estimated rates accounted fully for the measured overall rates. There were two striking results. First, the overall rates differed by an order of magnitude between substrates. Second, the relative contribution of each site was very different with different substrates. During succinate oxidation, most of the superoxide production was from the site of quinone reduction in complex I (site I Q), with small contributions from the flavin site in complex I (site I F) and the quinol oxidation site in complex III (site III Qo). However, with glutamate plus malate as substrate, site I Q made little or no contribution, and production was shared between site I F, site III Qo and 2-oxoglutarate dehydrogenase. With palmitoylcarnitine as substrate, the flavin site in complex II (site II F) was a major contributor (together with sites I F and III Qo), and with glycerol 3-phosphate as substrate, five different sites all contributed, including glycerol 3-phosphate dehydrogenase. Thus, the relative and absolute contributions of specific sites to the production of reactive oxygen species in isolated mitochondria depend very strongly on the substrates being oxidized, and the same is likely true in cells and in vivo.

          Graphical abstract

          Highlights

          • •

            Mitochondria oxidizing four substrate combinations make superoxide/H 2O 2 from six defined sites.

          • •

            The absolute rates of mitochondrial superoxide/H 2O 2 depend on the substrate oxidized.

          • •

            The relative contributions of specific sites of superoxide/H 2O 2 vary greatly between substrates.

          Related collections

          Most cited references28

          • Record: found
          • Abstract: found
          • Article: not found

          The sites and topology of mitochondrial superoxide production.

          Martin D Brand (2010)
          Mitochondrial superoxide production is an important source of reactive oxygen species in cells, and may cause or contribute to ageing and the diseases of ageing. Seven major sites of superoxide production in mammalian mitochondria are known and widely accepted. In descending order of maximum capacity they are the ubiquinone-binding sites in complex I (site IQ) and complex III (site IIIQo), glycerol 3-phosphate dehydrogenase, the flavin in complex I (site IF), the electron transferring flavoprotein:Q oxidoreductase (ETFQOR) of fatty acid beta-oxidation, and pyruvate and 2-oxoglutarate dehydrogenases. None of these sites is fully characterized and for some we only have sketchy information. The topology of the sites is important because it determines whether or not a site will produce superoxide in the mitochondrial matrix and be able to damage mitochondrial DNA. All sites produce superoxide in the matrix; site IIIQo and glycerol 3-phosphate dehydrogenase also produce superoxide to the intermembrane space. The relative contribution of each site to mitochondrial reactive oxygen species generation in the absence of electron transport inhibitors is unknown in isolated mitochondria, in cells or in vivo, and may vary considerably with species, tissue, substrate, energy demand and oxygen tension. Copyright (c) 2010 Elsevier Inc. All rights reserved.
          Article 0 comments Cited 347 times – based on 0 reviews     Review now
            Bookmark
            • Record: found
            • Abstract: found
            • Article: not found

            The cellular production of hydrogen peroxide.

            A Boveris, N Oshino, B. Chance (1972)
            1. The enzyme-substrate complex of yeast cytochrome c peroxidase is used as a sensitive, specific and accurate spectrophotometric H(2)O(2) indicator. 2. The cytochrome c peroxidase assay is suitable for use with subcellular fractions from tissue homogenates as well as with pure enzyme systems to measure H(2)O(2) generation. 3. Mitochondrial substrates entering the respiratory chain on the substrate side of the antimycin A-sensitive site support the mitochondrial generation of H(2)O(2). Succinate, the most effective substrate, yields H(2)O(2) at a rate of 0.5nmol/min per mg of protein in state 4. H(2)O(2) generation is decreased in the state 4-->state 3 transition. 4. In the combined mitochondrial-peroxisomal fraction of rat liver the changes in the mitochondrial generation of H(2)O(2) modulated by substrate, ADP and antimycin A are followed by parallel changes in the saturation of the intraperoxisomal catalase intermediate. 5. Peroxisomes supplemented with uric acid generate extraperoxisomal H(2)O(2) at a rate (8.6-16.4nmol/min per mg of protein) that corresponds to 42-61% of the rate of uric acid oxidation. Addition of azide increases these H(2)O(2) rates by a factor of 1.4-1.7. 6. The concentration of cytosolic uric acid is shown to vary during the isolation of the cellular fractions. 7. Microsomal fractions produce H(2)O(2) (up to 1.7nmol/min per mg of protein) at a ratio of 0.71-0.86mol of H(2)O(2)/mol of NADP(+) during the oxidation of NADPH. H(2)O(2) is also generated (6-25%) during the microsomal oxidation of NADH (0.06-0.025mol of H(2)O(2)/mol of NAD(+)). 8. Estimation of the rates of production of H(2)O(2) under physiological conditions can be made on the basis of the rates with the isolated fractions. The tentative value of 90nmol of H(2)O(2)/min per g of liver at 22 degrees C serves as a crude approximation to evaluate the biochemical impact of H(2)O(2) on cellular metabolism.
            Article 0 comments Cited 227 times – based on 0 reviews     Review now
              Bookmark
              • Record: found
              • Abstract: found
              • Article: not found

              Requirement for generation of H2O2 for platelet-derived growth factor signal transduction.

              M Sundaresan, Z X Yu, V Ferrans … (1995)
              Stimulation of rat vascular smooth muscle cells (VSMCs) by platelet-derived growth factor (PDGF) transiently increased the intracellular concentration of hydrogen peroxide (H2O2). This increase could be blunted by increasing the intracellular concentration of the scavenging enzyme catalase or by the chemical antioxidant N-acetylcysteine. The response of VSMCs to PDGF, which includes tyrosine phosphorylation, mitogen-activated protein kinase stimulation, DNA synthesis, and chemotaxis, was inhibited when the growth factor-stimulated rise in H2O2 concentration was blocked. These results suggest that H2O2 may act as a signal-transducing molecule, and they suggest a potential mechanism for the cardioprotective effects of antioxidants.
              Article 0 comments Cited 216 times – based on 0 reviews     Review now
                Bookmark
                All references

                Author and article information

                Contributors
                Casey L. Quinlan
                Journal
                Journal ID (nlm-ta): Redox Biol
                Journal ID (iso-abbrev): Redox Biol
                Title: Redox Biology
                Publisher: Elsevier
                ISSN (Electronic): 2213-2317
                Publication date PMC-release: 23 May 2013
                Publication date (Electronic): 23 May 2013
                Publication date Collection: 2013
                Volume: 1
                Issue: 1
                Pages: 304-312
                Affiliations
                [a ]The Buck Institute for Research on Aging, Novato, CA 94945, USA
                [b ]Department of Biomedical Sciences, Center for Healthy Aging, Copenhagen University, Denmark
                Author notes
                [* ]Correspondence to: The Buck Institute for Research on Aging, 8001 Redwood Blvd., Novato, CA 94945, USA. Tel.: +1 415 493 3676. cquinlan@ 123456buckinstitute.org
                Article
                Publisher ID: REDOX42
                DOI: 10.1016/j.redox.2013.04.005
                PMC ID: 3757699
                PubMed ID: 24024165
                SO-VID: 8694d5a3-b520-4448-a371-1fa35a9a4829
                Copyright © © 2013 The Authors
                License:

                This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-No Derivative Works License, which permits non-commercial use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                Date received : 22 March 2013
                Date revision received : 4 April 2013
                Date accepted : 5 April 2013
                Categories
                Subject: Research Paper

                Keywords: if, flavin site of complex i,iq, quinone-binding site of complex i,iif, flavin site of complex ii,iiiqo, quinol oxidation site of complex iii,cdnb, 1-chloro-2,4-dinitrobenzene,eh, redox potential,etf, electron transferring flavoprotein,etf:qor, etf:ubiquinone oxidoreductase,mgpdh, mitochondrial glycerol 3-phosphate dehydrogenase,ogdh, 2-oxoglutarate dehydrogenase,pdh, pyruvate dehydrogenase,q, ubiquinone,qh2, ubiquinol,ros, reactive oxygen species,superoxide,hydrogen peroxide,respiratory complexes,nadh,ubiquinone,cytochrome b
                Data availability:
                Keywords: if, flavin site of complex i, iq, quinone-binding site of complex i, iif, flavin site of complex ii, iiiqo, quinol oxidation site of complex iii, cdnb, 1-chloro-2,4-dinitrobenzene, eh, redox potential, etf, electron transferring flavoprotein, etf:qor, etf:ubiquinone oxidoreductase, mgpdh, mitochondrial glycerol 3-phosphate dehydrogenase, ogdh, 2-oxoglutarate dehydrogenase, pdh, pyruvate dehydrogenase, q, ubiquinone, qh2, ubiquinol, ros, reactive oxygen species, superoxide, hydrogen peroxide, respiratory complexes, nadh, ubiquinone, cytochrome b

                Comments

                Comment on this article