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Structural determinants of the high thermal stability of SsoPox from the hyperthermophilic archaeon Sulfolobus solfataricus

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Abstract

Organophosphates (OPs) constitute the largest class of insecticides used worldwide and certain of them are potent nerve agents. Consequently, enzymes degrading OPs are of paramount interest, as they could be used as bioscavengers and biodecontaminants. Looking for a stable OPs catalyst, able to support industrial process constraints, a hyperthermophilic phosphotriesterase (PTE) (SsoPox) was isolated from the archaeon Sulfolobus solfataricus and was found to be highly thermostable. The solved 3D structure revealed that SsoPox is a noncovalent dimer, with lactonase activity against “quorum sensing signals”, and therefore could represent also a potential weapon against certain pathogens. The structural basis of the high thermostability of SsoPox has been investigated by performing a careful comparison between its structure and that of two mesophilic PTEs from Pseudomonas diminuta and Agrobacterium radiobacter. In addition, the conformational stability of SsoPox against the denaturing action of temperature and GuHCl has been determined by means of circular dichroism and fluorescence measurements. The data suggest that the two fundamental differences between SsoPox and the mesophilic counterparts are: (a) a larger number of surface salt bridges, also involved in complex networks; (b) a tighter quaternary structure due to an optimization of the interactions at the interface between the two monomers.

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Abbreviations

OPs:

Organophosphates

PTE:

Phosphotriesterase

OPD:

Organophosphate-degrading

PLL:

Phosphotriesterase-like lactonase

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Acknowledgments

This research was supported by grants to E.C. by Délégation Générale pour l’Armement (CO no. 010807/03-10) and by the C.N.R.S. D.R. is under contract with Bundesministerium der Verteidigung (M/SAB/1/7/A004). We also thank MIUR project “Piano Nazionale Ricerca per le Biotecnologie Avanzate Tema II, Biocatalisi”.

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Authors and Affiliations

  1. Dipartimento di Chimica “Paolo Corradini”, Università di Napoli “Federico II”, Via Cintia, 80126, Naples, Italy

    Pompea Del Vecchio & Paola Carullo

  2. Architecture et Fonction des Macromolécules Biologiques, CNRS-Université de la Méditerranée, 13288, Marseille, France

    Mikael Elias & Eric Chabriere

  3. Istituto di Biochimica delle Proteine, Consiglio Nazionale delle Ricerche, Via P. Castellino 111, 80131, Naples, Italy

    Luigia Merone, Luigi Mandrich, Mosè Rossi & Giuseppe Manco

  4. Dipartimento di Scienze Biologiche ed Ambientali, Università del Sannio, 82100, Benevento, Italy

    Giuseppe Graziano

  5. Laboratoire de Cristallogenèse et Cristallographie des Protéines, Institut de Biologie Structurale JP EBEL, 38027, Grenoble, France

    Jérôme Dupuy

  6. Laboratoire de Cristallographie et Modélisation des Matériaux Minéraux et Biologiques, CNRS-Université Henri Poincaré, 54506, Nancy, France

    Bertrand Fournier

  7. Unité d’Enzymologie, Département de Toxicologie, Centre de Recherches du Service de Santé des Armées, 38702, La Tronche, France

    Daniel Rochu, Patrick Masson & Eric Chabriere

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  1. Pompea Del Vecchio
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  2. Mikael Elias
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  3. Luigia Merone
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  4. Giuseppe Graziano
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  5. Jérôme Dupuy
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  6. Luigi Mandrich
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  7. Paola Carullo
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  9. Daniel Rochu
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  10. Mosè Rossi
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  11. Patrick Masson
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  13. Giuseppe Manco
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Corresponding author

Correspondence to Giuseppe Manco.

Additional information

Communicated by T. Matsunaga.

Pompea Del Vecchio, Mikael Elias and Luigia Merone were contributed equally to this paper.

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Del Vecchio, P., Elias, M., Merone, L. et al. Structural determinants of the high thermal stability of SsoPox from the hyperthermophilic archaeon Sulfolobus solfataricus . Extremophiles 13, 461–470 (2009). https://doi.org/10.1007/s00792-009-0231-9

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