Reduction of the Iron−Sulfur Clusters in Mitochondrial NADH:Ubiquinone Oxidoreductase (Complex I) by EuII-DTPA, a Very Low Potential Reductant† (original) (raw)

ArticleJuly 24, 2008

Biochemistry

Cite this: Biochemistry 2008, 47, 34

Click to copy citationCitation copied!

research-article

Copyright © 2008 American Chemical Society

Abstract

Click to copy section linkSection link copied!

Abstract Image

NADH:ubiquinone oxidoreductase (complex I) is the first enzyme of the mitochondrial electron transport chain. It contains a flavin mononucleotide to oxidize NADH, and eight iron-sulfur clusters. Seven of them transfer electrons between the flavin and the quinone-binding site, and one is on the opposite side of the flavin. Although most information about their properties is from EPR, the spectra from only five clusters have been observed, and it is difficult to match them to the structurally defined clusters. Here, we analyze complex I from bovine mitochondria reacted with a very low potential reductant, to impose a potential approaching −1 V. We compare the spectra with those from higher potentials and from the 24 kDa subunit and flavoprotein subcomplex, and model the spectra by starting from those with fewer components and building the complexity gradually. Spectrum N1a, from the 24 kDa subunit [2Fe-2S] cluster, is not observed in bovine complex I at any potential. Spectrum N1b, from the 75 kDa subunit [2Fe-2S] cluster, exhibits a lower potential than the N3, N4 and N5 spectra of three [4Fe-4S] clusters. In the lowest potential spectra an N5-type spectrum is observed at unusually high temperature (indicating a significant change to the cluster, or that two clusters have very similar g values), the relaxation rate of N1b increases (indicating that a nearby cluster has become reduced) and a new feature with an apparent g value of 2.16 suggests an interaction between two reduced clusters. The consequences of these observations for electron transfer in complex I are discussed.

ACS Publications

Copyright © 2008 American Chemical Society

Supporting Information

Click to copy section linkSection link copied!

Comparison of the principal g values of modeled spectra with their published equivalents; spectra recorded over a range of temperature for each sample analyzed; wide-field spectrum of EuII sample. This material is available free of charge via the Internet at http://pubs.acs.org.

Terms & Conditions

Most electronic Supporting Information files are available without a subscription to ACS Web Editions. Such files may be downloaded by article for research use (if there is a public use license linked to the relevant article, that license may permit other uses). Permission may be obtained from ACS for other uses through requests via the RightsLink permission system: http://pubs.acs.org/page/copyright/permissions.html.

Cited By

Click to copy section linkSection link copied!

This article is cited by 47 publications.

  1. Frederic Melin, Petra Hellwig. Redox Properties of the Membrane Proteins from the Respiratory Chain. Chemical Reviews 2020, 120 (18) , 10244-10297. https://doi.org/10.1021/acs.chemrev.0c00249
  2. Tyler C. Jenks, Akhila N. W. Kuda-Wedagedara, Matthew D. Bailey, Cassandra L. Ward, Matthew J. Allen. Spectroscopic and Electrochemical Trends in Divalent Lanthanides through Modulation of Coordination Environment. Inorganic Chemistry 2020, 59 (4) , 2613-2620. https://doi.org/10.1021/acs.inorgchem.0c00136
  3. Katerina Dörner, Marta Vranas, Johannes Schimpf, Isabella R. Straub, Jo Hoeser, and Thorsten Friedrich . Significance of [2Fe-2S] Cluster N1a for Electron Transfer and Assembly of Escherichia coli Respiratory Complex I. Biochemistry 2017, 56 (22) , 2770-2778. https://doi.org/10.1021/acs.biochem.6b01058
  4. Maxie M. Roessler, Rhiannon M. Evans, Rosalind A. Davies, Jeffrey Harmer, and Fraser A. Armstrong . EPR Spectroscopic Studies of the Fe–S Clusters in the O2-Tolerant [NiFe]-Hydrogenase Hyd-1 from Escherichia coli and Characterization of the Unique [4Fe–3S] Cluster by HYSCORE. Journal of the American Chemical Society 2012, 134 (37) , 15581-15594. https://doi.org/10.1021/ja307117y
  5. Hannah R. Bridges, Eckhard Bill, and Judy Hirst . Mössbauer Spectroscopy on Respiratory Complex I: The Iron–Sulfur Cluster Ensemble in the NADH-Reduced Enzyme Is Partially Oxidized. Biochemistry 2012, 51 (1) , 149-158. https://doi.org/10.1021/bi201644x
  6. Martin S. King, Mark S. Sharpley and Judy Hirst . Reduction of Hydrophilic Ubiquinones by the Flavin in Mitochondrial NADH:Ubiquinone Oxidoreductase (Complex I) and Production of Reactive Oxygen Species. Biochemistry 2009, 48 (9) , 2053-2062. https://doi.org/10.1021/bi802282h
  7. Daniel Wohlwend, Thilo Seifermann, Emmanuel Gnandt, Marta Vranas, Stefan Gerhardt, Thorsten Friedrich. Structural changes shifting the redox potential of the outlying cluster N1a in respiratory complex I. Structure 2026, 34 (1) , 175-183.e3. https://doi.org/10.1016/j.str.2025.10.016
  8. Wilfred R. Hagen. The g-Strained EPR Line Shape of Transition-Ion Complexes and Metalloproteins: Four Decades of Misunderstanding and Its Consequences. Molecules 2025, 30 (15) , 3299. https://doi.org/10.3390/molecules30153299
  9. Anjali Depala, Manon T. Lachmann, Simone Morra, James A. Birrell, Patricia Rodríguez-Maciá. Terminal hydride formation in [FeFe] hydrogenase: understanding the role of the dithiolate bridgehead. Chemical Communications 2025, 61 (33) , 6178-6181. https://doi.org/10.1039/D5CC00860C
  10. Elisabeth Lettau, Christian Lorent, Jens Appel, Marko Boehm, Paul R.F. Cordero, Lars Lauterbach. Insights into electron transfer and bifurcation of the Synechocystis sp. PCC6803 hydrogenase reductase module. Biochimica et Biophysica Acta (BBA) - Bioenergetics 2025, 1866 (1) , 149508. https://doi.org/10.1016/j.bbabio.2024.149508
  11. Vera G. Grivennikova, Grigory V. Gladyshev, Tatyana V. Zharova, Vitaliy B. Borisov. Proton-Translocating NADH–Ubiquinone Oxidoreductase: Interaction with Artificial Electron Acceptors, Inhibitors, and Potential Medicines. International Journal of Molecular Sciences 2024, 25 (24) , 13421. https://doi.org/10.3390/ijms252413421
  12. Benjamin R. Duffus, Marcel Gauglitz, Christian Teutloff, Silke Leimkühler. Redox potentials elucidate the electron transfer pathway of NAD+-dependent formate dehydrogenases. Journal of Inorganic Biochemistry 2024, 253 , 112487. https://doi.org/10.1016/j.jinorgbio.2024.112487
  13. Rikuan Zheng, Chong Wang, Rui Liu, Ruining Cai, Chaomin Sun. Physiological and metabolic insights into the first cultured anaerobic representative of deep-sea Planctomycetes bacteria. eLife 2024, 12 https://doi.org/10.7554/eLife.89874
  14. Rikuan Zheng, Chong Wang, Rui Liu, Ruining Cai, Chaomin Sun. Physiological and metabolic insights into the first cultured anaerobic representative of deep-sea Planctomycetes bacteria. eLife 2024, 12 https://doi.org/10.7554/eLife.89874.3
  15. Owen D. Jarman, Olivier Biner, John J. Wright, Judy Hirst. Paracoccus denitrificans: a genetically tractable model system for studying respiratory complex I. Scientific Reports 2021, 11 (1)https://doi.org/10.1038/s41598-021-89575-9
  16. Hannah R. Bridges, Justin G. Fedor, James N. Blaza, Andrea Di Luca, Alexander Jussupow, Owen D. Jarman, John J. Wright, Ahmed-Noor A. Agip, Ana P. Gamiz-Hernandez, Maxie M. Roessler, Ville R. I. Kaila, Judy Hirst. Structure of inhibitor-bound mammalian complex I. Nature Communications 2020, 11 (1)https://doi.org/10.1038/s41467-020-18950-3
  17. Russ Hille, Tynan Young, Dimitri Niks, Sheron Hakopian, Timothy K. Tam, Xuejun Yu, Ashok Mulchandani, Gregor M. Blaha. Structure: Function Studies of the Cytosolic, Mo- and NAD+-Dependent Formate Dehydrogenase from Cupriavidus necator. Inorganics 2020, 8 (7) , 41. https://doi.org/10.3390/inorganics8070041
  18. Tynan Young, Dimitri Niks, Sheron Hakopian, Timothy K. Tam, Xuejun Yu, Russ Hille, Gregor M. Blaha. Crystallographic and kinetic analyses of the FdsBG subcomplex of the cytosolic formate dehydrogenase FdsABG from Cupriavidus necator. Journal of Biological Chemistry 2020, 295 (19) , 6570-6585. https://doi.org/10.1074/jbc.RA120.013264
  19. Marius Schulte, Klaudia Frick, Emmanuel Gnandt, Sascha Jurkovic, Sabrina Burschel, Ramona Labatzke, Karoline Aierstock, Dennis Fiegen, Daniel Wohlwend, Stefan Gerhardt, Oliver Einsle, Thorsten Friedrich. A mechanism to prevent production of reactive oxygen species by Escherichia coli respiratory complex I. Nature Communications 2019, 10 (1)https://doi.org/10.1038/s41467-019-10429-0
  20. Hendrike Dürichen, Gabriele Diekert, Sandra Studenik. Redox potential changes during ATP‐dependent corrinoid reduction determined by redox titrations with europium(II)–DTPA. Protein Science 2019, 28 (10) , 1902-1908. https://doi.org/10.1002/pro.3699
  21. Kelly N. Tran, Shuqiang Niu, Toshiko Ichiye. Reduction potential calculations of the Fe–S clusters in Thermus thermophilus respiratory complex I. Journal of Computational Chemistry 2019, 40 (12) , 1248-1256. https://doi.org/10.1002/jcc.25785
  22. William E. Antholine, Jeannette Vasquez-Vivar, Brendan J. Quirk, Harry T. Whelan, Pui Kei Wu, Jong-In Park, Charles R. Myers. Treatment of Cells and Tissues with Chromate Maximizes Mitochondrial 2Fe2S EPR Signals. International Journal of Molecular Sciences 2019, 20 (5) , 1143. https://doi.org/10.3390/ijms20051143
  23. Jan M. Schuller, James A. Birrell, Hideaki Tanaka, Tsuyoshi Konuma, Hannes Wulfhorst, Nicholas Cox, Sandra K. Schuller, Jacqueline Thiemann, Wolfgang Lubitz, Pierre Sétif, Takahisa Ikegami, Benjamin D. Engel, Genji Kurisu, Marc M. Nowaczyk. Structural adaptations of photosynthetic complex I enable ferredoxin-dependent electron transfer. Science 2019, 363 (6424) , 257-260. https://doi.org/10.1126/science.aau3613
  24. M. Mervan Çakar, Juan Mangas-Sanchez, William R. Birmingham, Nicholas J. Turner, Barış Binay. Discovery of a new metal and NAD + -dependent formate dehydrogenase from Clostridium ljungdahlii. Preparative Biochemistry & Biotechnology 2018, 48 (4) , 327-334. https://doi.org/10.1080/10826068.2018.1446150
  25. Janina Preissler, Stefan Wahlefeld, Christian Lorent, Christian Teutloff, Marius Horch, Lars Lauterbach, Stephen P. Cramer, Ingo Zebger, Oliver Lenz. Enzymatic and spectroscopic properties of a thermostable [NiFe]‑hydrogenase performing H2-driven NAD+-reduction in the presence of O2. Biochimica et Biophysica Acta (BBA) - Bioenergetics 2018, 1859 (1) , 8-18. https://doi.org/10.1016/j.bbabio.2017.09.006
  26. Emmanuel Gnandt, Johannes Schimpf, Caroline Harter, Jo Hoeser, Thorsten Friedrich. Reduction of the off-pathway iron-sulphur cluster N1a of Escherichia coli respiratory complex I restrains NAD+ dissociation. Scientific Reports 2017, 7 (1)https://doi.org/10.1038/s41598-017-09345-4
  27. John J. Wright, Enrico Salvadori, Hannah R. Bridges, Judy Hirst, Maxie M. Roessler. Small-volume potentiometric titrations: EPR investigations of Fe-S cluster N2 in mitochondrial complex I. Journal of Inorganic Biochemistry 2016, 162 , 201-206. https://doi.org/10.1016/j.jinorgbio.2016.04.025
  28. Judy Hirst, Maxie M. Roessler. Energy conversion, redox catalysis and generation of reactive oxygen species by respiratory complex I. Biochimica et Biophysica Acta (BBA) - Bioenergetics 2016, 1857 (7) , 872-883. https://doi.org/10.1016/j.bbabio.2015.12.009
  29. Hannah R. Bridges, Andrew J. Y. Jones, Michael N. Pollak, Judy Hirst. Effects of metformin and other biguanides on oxidative phosphorylation in mitochondria. Biochemical Journal 2014, 462 (3) , 475-487. https://doi.org/10.1042/BJ20140620
  30. John M Berrisford, Rozbeh Baradaran, Leonid A Sazanov. Entire Respiratory Complex I from Thermus Thermophilus. 2014, 1-16. https://doi.org/10.1002/9781119951438.eibc2266
  31. James A. Birrell, Klaudia Morina, Hannah R. Bridges, Thorsten Friedrich, Judy Hirst. Investigating the function of [2Fe–2S] cluster N1a, the off-pathway cluster in complex I, by manipulating its reduction potential. Biochemical Journal 2013, 456 (1) , 139-146. https://doi.org/10.1042/BJ20130606
  32. Judy Hirst. Mitochondrial Complex I. Annual Review of Biochemistry 2013, 82 (1) , 551-575. https://doi.org/10.1146/annurev-biochem-070511-103700
  33. Bradley Scott Perrin, Shuqiang Niu, Toshiko Ichiye. Calculating standard reduction potentials of [4Fe–4S] proteins. Journal of Computational Chemistry 2013, 34 (7) , 576-582. https://doi.org/10.1002/jcc.23169
  34. Marina Verkhovskaya, Dmitry A. Bloch. Energy-converting respiratory Complex I: On the way to the molecular mechanism of the proton pump. The International Journal of Biochemistry & Cell Biology 2013, 45 (2) , 491-511. https://doi.org/10.1016/j.biocel.2012.08.024
  35. M. Horch, L. Lauterbach, O. Lenz, P. Hildebrandt, I. Zebger. NAD(H)‐coupled hydrogen cycling – structure–function relationships of bidirectional [NiFe] hydrogenases. FEBS Letters 2012, 586 (5) , 545-556. https://doi.org/10.1016/j.febslet.2011.10.010
  36. Eiko Nakamaru-Ogiso. Iron–Sulfur Clusters in Complex I. 2012, 61-79. https://doi.org/10.1007/978-94-007-4138-6_3
  37. Leonid Sazanov. Hydrophilic Domain of Respiratory Complex I from Thermus thermophilus. 2011https://doi.org/10.1002/9781119951438.eibc0666
  38. Kenneth R. Pryde, Judy Hirst. Superoxide Is Produced by the Reduced Flavin in Mitochondrial Complex I. Journal of Biological Chemistry 2011, 286 (20) , 18056-18065. https://doi.org/10.1074/jbc.M110.186841
  39. Charles R. Myers, William E. Antholine, Judith M. Myers. The pro-oxidant chromium(VI) inhibits mitochondrial complex I, complex II, and aconitase in the bronchial epithelium: EPR markers for Fe–S proteins. Free Radical Biology and Medicine 2010, 49 (12) , 1903-1915. https://doi.org/10.1016/j.freeradbiomed.2010.09.020
  40. Emile S. Medvedev, Vernon A. Couch, Alexei A. Stuchebrukhov. Determination of the intrinsic redox potentials of FeS centers of respiratory complex I from experimental titration curves. Biochimica et Biophysica Acta (BBA) - Bioenergetics 2010, 1797 (9) , 1665-1671. https://doi.org/10.1016/j.bbabio.2010.05.011
  41. Simon P. J. Albracht. The reaction of NADPH with bovine mitochondrial NADH:ubiquinone oxidoreductase revisited. Journal of Bioenergetics and Biomembranes 2010, 42 (4) , 261-278. https://doi.org/10.1007/s10863-010-9301-z
  42. Simon P. J. Albracht. The reaction of NADPH with bovine mitochondrial NADH:ubiquinone oxidoreductase revisited. Journal of Bioenergetics and Biomembranes 2010, 42 (4) , 279-292. https://doi.org/10.1007/s10863-010-9302-y
  43. Maxie M. Roessler, Martin S. King, Alan J. Robinson, Fraser A. Armstrong, Jeffrey Harmer, Judy Hirst. Direct assignment of EPR spectra to structurally defined iron-sulfur clusters in complex I by double electron–electron resonance. Proceedings of the National Academy of Sciences 2010, 107 (5) , 1930-1935. https://doi.org/10.1073/pnas.0908050107
  44. Judy Hirst. Towards the molecular mechanism of respiratory complex I. Biochemical Journal 2010, 425 (2) , 327-339. https://doi.org/10.1042/BJ20091382
  45. Hannah R. Bridges, Ljuban Grgic, Michael E. Harbour, Judy Hirst. The respiratory complexes I from the mitochondria of two Pichia species. Biochemical Journal 2009, 422 (1) , 151-159. https://doi.org/10.1042/BJ20090492
  46. Volker Zickermann, Stefan Kerscher, Klaus Zwicker, Maja A. Tocilescu, Michael Radermacher, Ulrich Brandt. Architecture of complex I and its implications for electron transfer and proton pumping. Biochimica et Biophysica Acta (BBA) - Bioenergetics 2009, 1787 (6) , 574-583. https://doi.org/10.1016/j.bbabio.2009.01.012
  47. Leonid Sazanov. Hydrophilic Domain of Respiratory Complex I from Thermus thermophilus. 2004https://doi.org/10.1002/0470028637.met244

Biochemistry

Cite this: Biochemistry 2008, 47, 34

Click to copy citationCitation copied!

Copyright © 2008 American Chemical Society

Altmetric

-

Citations

Learn about these metrics

Article Views are the COUNTER-compliant sum of full text article downloads since November 2008 (both PDF and HTML) across all institutions and individuals. These metrics are regularly updated to reflect usage leading up to the last few days.

Citations are the number of other articles citing this article, calculated by Crossref and updated daily. Find more information about Crossref citation counts.

The Altmetric Attention Score is a quantitative measure of the attention that a research article has received online. Clicking on the donut icon will load a page at altmetric.com with additional details about the score and the social media presence for the given article. Find more information on the Altmetric Attention Score and how the score is calculated.