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Cloning, expression and physicochemical characterization of a di-heme cytochrome c 4 from the psychrophilic bacterium Pseudoalteromonas haloplanktis TAC 125

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Abstract

The 20-kDa di-heme cytochrome c 4 from the psycrophilic bacterium Pseudoalteromonas haloplanktis TAC 125 was cloned and expressed in Escherichia coli and investigated through UV–vis and 1H NMR spectroscopies and protein voltammetry. The model structure was computed using the X-ray structure of Pseudomonas stutzeri cytochrome c 4 as a template. The protein shows unprecedented properties within the cytochrome c 4 family, including (1) an almost nonpolar surface charge distribution, (2) the absence of high-spin heme Fe(III) states, indicative of a thermodynamically stable and kinetically inert axial heme His,Met coordination, and (3) identical E°′ values for the two heme centers (+0.322 V vs the standard hydrogen elecrode). At pH extremes, both heme groups undergo the “acid” and “alkaline” conformational transitions typical of class I cytochromes c, involving ligand-exchange equilibria, whereas at intermediate pH values their electronic properties are sensitive to several residue ionizations.

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Acknowledgments

This work was supported by the University of Modena and Reggio Emilia (Fondo di Ateneo per la Ricerca, 2006–2007). NAMD was developed by the Theoretical and Computational Biophysics Group in the Beckman Institute for Advanced Science and Technology at the University of Illinois at Urbana-Champaign.

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Authors and Affiliations

  1. Department of Chemistry, Università di Modena and Reggio Emilia, Via Campi 183, 41100, Modena, Italy

    Giulia Di Rocco, Gianantonio Battistuzzi, Marco Borsari, Francesca De Rienzo, Antonio Ranieri & Marco Sola

  2. CNR-INFM National Center nanoStructures and bioSystems at Surfaces, S3, Via Campi 213/A, 41100, Modena, Italy

    Francesca De Rienzo & Marco Sola

  3. Dipartimento di Chimica Organica e Biochimica, Università di Napoli “Federico II”, Monte Sant’Angelo, Via Cinthia 4, 80126, Naples, Italy

    Maria Luisa Tutino

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  1. Giulia Di Rocco
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  2. Gianantonio Battistuzzi
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  4. Francesca De Rienzo
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  6. Maria Luisa Tutino
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  7. Marco Sola
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Correspondence to Marco Sola.

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Di Rocco, G., Battistuzzi, G., Borsari, M. et al. Cloning, expression and physicochemical characterization of a di-heme cytochrome c 4 from the psychrophilic bacterium Pseudoalteromonas haloplanktis TAC 125. J Biol Inorg Chem 13, 789–799 (2008). https://doi.org/10.1007/s00775-008-0366-7

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  • DOI: https://doi.org/10.1007/s00775-008-0366-7

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