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Kinetics of the electron transfer reaction of Cytochrome c 552 adsorbed on biomimetic electrode studied by time-resolved surface-enhanced resonance Raman spectroscopy and electrochemistry

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Abstract

Cytochrome c 552 (Cyt-c 552) and its redox partner ba 3 -oxidase from Thermus thermophilus possess structural differences compared with Horse heart cytochrome c (cyt-c)/cytochrome c oxidase (CcO) system, where the recognition between partners and the electron transfer (ET) process is initiated via electrostatic interactions. We demonstrated in a previous study by surface-enhanced resonance Raman (SERR) spectroscopy that roughened silver electrodes coated with uncharged mixed self-assembled monolayers HS–(CH2) n –CH3/HS–(CH2) n + 1–OH 50/50, n = 5, 10 or 15, was a good model to mimic the Cyt-c 552 redox partner. All the adsorbed molecules are well oriented on such biomimetic electrodes and transfer one electron during the redox process. The present work focuses on the kinetic part of the heterogeneous ET process of Cyt-c 552 adsorbed onto electrodes coated with such mixed SAMs of different alkyl chain length. For that purpose, two complementary methods were combined. Firstly cyclic voltammetry shows that the ET between the adsorbed Cyt-c 552 and the biomimetic electrode is direct and reversible. Furthermore, it allows the estimation of both the density surface coverage of adsorbed Cyt-c 552 and the kinetic constants values. Secondly, time-resolved SERR (TR-SERR) spectroscopy showed that the ET process occurs without conformational change of the Cyt-c 552 heme group and allows the determination of kinetic constants. Results show that the kinetic constant values obtained by TR-SERR spectroscopy could be compared to those obtained from cyclic voltammetry. They are estimated at 200, 150 and 40 s−1 for the ET of Cyt-c 552 adsorbed onto electrodes coated with mixed SAMs HS–(CH2) n –CH3/HS–(CH2) n + 1–OH 50/50, n = 5, 10 or 15, respectively.

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References

  • Avila A, Gregory BW, Niki K, Cotton TM (2000) An electrochemical approach to investigate gated electron transfer using a physiological model system: cytochrome c immobilized on carboxylic acid-terminated alkanethiol self-assembled monolayers on gold electrodes. J Phys Chem B 104:2759–2766

    Article  Google Scholar 

  • Bard AJ, Faulkner LR (2001) Electrochemical methods: fundamentals and applications. Wiley, New York

    Google Scholar 

  • Battistuzzi G, Borsari M, Cowan JA, Ranieri A, Sola M (2002) Control of cytochrome c redox potential: axial ligation and protein environment effect. J Am Chem Soc 124:5315–5324

    Article  Google Scholar 

  • Bernad S, Soulimane T, Lecomte S (2004) Redox and conformational equilibria of cytochrome c552 from Thermus thermipohilus adsorbed on chemically modified silver electrode probed by SERR spectroscopy. J Raman Spectrosc 35:47–54

    Article  Google Scholar 

  • Bernad S, Soulimane T, Lecomte S (2006) Characterization and redox properties of cytochrome c552 from Thermus thermophilus adsorbed on different self-assembled thiol monolayers, used to model the chemical environment of the redox partner. Biopolymers 81:407–418

    Article  Google Scholar 

  • Clark RA, Bowden EF (1997) Voltametric peak broadening for cytochrome c/alkanethiolate monolayer structures: dispersion of formal potentials. Langmuir 13:559–565

    Article  Google Scholar 

  • Cotton TM, Schultz SG, Van Duyne RP (1980) Surface-enhanced resonance Raman scattering from cytochrome c and myoglobin adsorbed on a silver electrode. J Am Chem Soc 102:7960–7962

    Article  Google Scholar 

  • Döpner S, Hildebrandt P, Rosell FI, Mauk AG, von Walter M, Buse G, Soulimane T (1999) The structural and functional role of lysine residues in the binding domain of cytochrome c in the electron transfer to cytochrome c oxidase. Eur J Biochem 261:379–391

    Article  Google Scholar 

  • El Kasmi A, Wallace JM, Bowden EF, Binet SM, Linderman RJ (1998) Controlling interfacial electron transfer kinetics of cytochrome c with mixed self-assembled monolayers. J Am Chem Soc 120:225–226

    Article  Google Scholar 

  • Feng ZQ, Imabayashi S, T. K, Niki K (1995) Electroreflectance spectroscopic study of the electron transfer rate of cytochrome c electrostatically immobilized on the o-carboxyl alkanethiol monolayer modified gold electrode. J Electroanal Chem 394:149–154

  • Feng ZQ, Imabayashi S, Kakiushi T, Niki K (1997) Long-range electron-transfer reaction rates to cytochrome across long- and short- chain alkanethiol self-assembled monolayers: electroreflectance studies. J Chem Soc Faraday Trans 93:1367–1370

    Article  Google Scholar 

  • Hildebrandt P, Macor KA, Czernuszewicz RS (1988) Novel cylindrical rotating electrode for anaerobic surface-enhanced Raman spectroscopy. J Raman Spectrosc 19:65–69

    Article  Google Scholar 

  • Hildebrandt P, Murgida DH (2002) Electron transfer dynamics of cytochrome c bound to self-assembled monolayers on silver electrodes. Bioelectrochemistry 55:139–143

    Article  Google Scholar 

  • Hildebrandt P, Stockburger M (1989a) Cytochrome c at charged interfaces 1: conformational and redox equilibria at the electrode/electrolyte interface probed by surface-enhanced resonance Raman spectroscopy. Biochemistry 28:6710–6721

    Article  Google Scholar 

  • Hildebrandt P, Stockburger M (1989b) Cytochrome c at charged interfaces 2: complexes with negatively charged macromolecular systems studied by resonance Raman spectroscopy. Biochemistry 28:6722–6728

    Article  Google Scholar 

  • Hon-Nami K, Oshim T (1977) Purification and some properties of cytochrome c552 from an extreme thermophile, Thermus thermophilus. J Biochem 82:769–776

    Google Scholar 

  • Hu S, Morris IK, Singh JP, Smith KM, Spiro TG (1993) Complete assignment of cytochrome c resonance Raman spectra via enzymatic reconstitution with isotopically labeled hemes. J Am Chem Soc 115:12446–12458

    Article  Google Scholar 

  • Jeuken LJC (2003) Conformational reorganisation in interfacial protein electron transfer. Biochim Biophys Acta 1604:67–76

    Article  Google Scholar 

  • Laviron E (1979) General expression of the linear potential sweep voltammogram in the case of diffusionless electrochemical systems. J Electroanal Chem 101:19–28

    Article  Google Scholar 

  • Lecomte S, Hildebrandt P, Soulimane T (1999) Dynamics of the heterogeneous electron-transfer reaction of cytochrome c 552 from Thermus thermophilus. A time-resolved surface-enhanced resonance Raman spectroscopic study. J Phys Chem B 103:10053–10064

    Article  Google Scholar 

  • Lecomte S, Wackerbarth H, Hildebrandt P, Soulimane T, Buse G (1998a) Potential-dependent surface-enhanced resonance Raman spectroscopy of cytochrome c 552 from Thermus thermophilus. J Raman Spectrosc 29:687–692

    Article  Google Scholar 

  • Lecomte S, Wackerbarth H, Soulimane T, Buse G, Hildebrandt P (1998b) Time-resolved surface-enhanced resonance Raman spectroscopy for stuying electron -transfer dynamics of heme proteins. J Am Chem Soc 120:7381–7382

    Article  Google Scholar 

  • Leopold MC, Bowden EF (2002) Influence of gold substrate topography on the voltammetry of cytochrome c adsorbed on carboxylic acid terminated self-assembled monolayers. Langmuir 18:2239–2245

    Article  Google Scholar 

  • Marcus RA, Sutin N (1985) Electron transfers in chemistry and biology. Biochim Biophys Acta 811:265–322

    Google Scholar 

  • Murgida DH, Hildebrandt P (2001) Proton-coupled electron transfer of cytochrome c. J Am Chem Soc 123:4062–4068

    Article  Google Scholar 

  • Murgida DH, Hildebrandt P (2004) Electron-transfer processes of cytochrome c at interfaces: new insights by surface-enhanced resonance Raman spectroscopy. Acc Chem Res 37:854–861

    Article  Google Scholar 

  • Scott RA, Mauk AG (1995) Cytochrome c: a multidisciplinary approach. University Science Books, Sausalito

    Google Scholar 

  • Song S, Clark RA, Bowden EF (1993) Characterization of cytochrome c/alkanethiolate structures prepared by self-assembly on gold. J Phys Chem 97:6564–6572

    Article  Google Scholar 

  • Soulimane T, Buse G, Bourenkov GP, Bartunik HD, Huber R, Than ME (2000) Structure and mechanism of the aberrant ba 3-cytochrome c oxydases from Thermus thermophilus. EMBO Journal 19:1766–1776

    Article  Google Scholar 

  • Soulimane T, Von Walter M, Hof P, Than ME, Huber R, Buse G (1997) Cytochrome c 552 from Thermus thermophilus: a functional and crystallographic investigation. Biochemical and Biophysical Research Commun 237:572–576

    Article  Google Scholar 

  • Than ME, Hof P, Huber R, Bourenkov GP, Bartunik HD, Buse G, Soulimane T (1997) Thermus thermophilus cytochrome c 552: a new highly thermostable cytochrome c structure obtained by MAD phasing. J Mol Biol 271:629–644

    Article  Google Scholar 

  • Than ME, Soulimane T (2001) Ba 3-Cytochrome c oxydase from Thermus thermophilus. In: Messerschmidt A, Huber R, Poulos T, Wieghardt K (eds) Handbook of metalloproteins, Chichester, pp 363–378

  • Wackerbarth H, Klar U, Günther W, Hildebrandt P (1999) Novel time-resolved surface-enhanced (resonance) Raman spectroscopic technique for studying the dynamics of interfacial processes: application to the electron transfer reaction of cytochrome c at silver electrode. Appl Spectrosc 53:283–291

    Article  ADS  Google Scholar 

  • Wei J, Liu AR, Yamamoto H, He Y, Waldeck DH (2002) Direct wiring of cytochrme c’s heme unit to an electrode: electrochemical studies. J Am Chem Soc 124:9591–9599

    Article  Google Scholar 

  • Wei JJ, Liu H, Niki K, Margoliash E, Waldeck DH (2004) Probing electron tunneling pathways: electrochemical study of rat heart cytochrome c and its mutant on pyridine-terminated SAMs. J Phys Chem B 108:16912–16917

    Article  Google Scholar 

  • Yue H, Khoshtariya D, Waldeck DH, Grochol J, Hildebrandt P, Murgida DH (2006) On the electron transfer mechanism between cytochrome c and metal electrodes: evidence for dynamics control at short distances. J Phys Chem B 110:19906–19913

    Article  Google Scholar 

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Acknowledgment

The authors thank Dr Tewfik Soulimane for the gift of Cyt-c 552.

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Authors and Affiliations

  1. LADIR, CNRS/UPMC (UMR 7075), 2 rue Henri Dunant, 94320, Thiais, France

    Sophie Bernad, Nadine Leygue & Sophie Lecomte

  2. Equipe de Chimie Bioorganique et Bioinorganique, Institut de Chimie Moléculaire et des Matériaux d’Orsay, UMR-CNRS 8182, Bâtiment 420, université Paris-Sud XI, 91405, Orsay, France

    Hafsa Korri-Youssoufi

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  1. Sophie Bernad
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  2. Nadine Leygue
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  3. Hafsa Korri-Youssoufi
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  4. Sophie Lecomte
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Correspondence to Sophie Lecomte.

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Bernad, S., Leygue, N., Korri-Youssoufi, H. et al. Kinetics of the electron transfer reaction of Cytochrome c 552 adsorbed on biomimetic electrode studied by time-resolved surface-enhanced resonance Raman spectroscopy and electrochemistry. Eur Biophys J 36, 1039–1048 (2007). https://doi.org/10.1007/s00249-007-0173-z

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  • DOI: https://doi.org/10.1007/s00249-007-0173-z

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