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

Pompea Del Vecchio; Mikael Elias; Luigia Merone; Giuseppe Graziano; Jérôme Dupuy; Luigi Mandrich; Paola Carullo; Bertrand Fournier; Daniel Rochu; Mosè Rossi; Patrick Masson; Eric Chabriere; Giuseppe Manco

doi:10.1007/s00792-009-0231-9

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

Pompea Del Vecchio, Mikael Elias, Luigia Merone, Giuseppe Graziano, Jérôme Dupuy, Luigi Mandrich, Paola Carullo, Bertrand Fournier, Daniel Rochu, Mosè Rossi, Patrick Masson, Eric Chabriere, Giuseppe Manco

Biochemistry, Molecular Biology, and Biophysics (CBS)

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53 Scopus citations

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.

Original language	English (US)
Pages (from-to)	461-470
Number of pages	10
Journal	Extremophiles
Volume	13
Issue number	3
DOIs	https://doi.org/10.1007/s00792-009-0231-9
State	Published - May 2009

Bibliographical note

Funding Information:
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’’.

Keywords

Conformational stability
Hyperthermophilic enzyme
Quaternary structure organization
Salt bridges

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10.1007/s00792-009-0231-9

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Del Vecchio, P., Elias, M., Merone, L., Graziano, G., Dupuy, J., Mandrich, L., Carullo, P., Fournier, B., Rochu, D., Rossi, M., Masson, P., Chabriere, E., & Manco, G. (2009). Structural determinants of the high thermal stability of SsoPox from the hyperthermophilic archaeon Sulfolobus solfataricus. Extremophiles, 13(3), 461-470. https://doi.org/10.1007/s00792-009-0231-9

Del Vecchio, P, Elias, M, Merone, L, Graziano, G, Dupuy, J, Mandrich, L, Carullo, P, Fournier, B, Rochu, D, Rossi, M, Masson, P, Chabriere, E & Manco, G 2009, 'Structural determinants of the high thermal stability of SsoPox from the hyperthermophilic archaeon Sulfolobus solfataricus', Extremophiles, vol. 13, no. 3, pp. 461-470. https://doi.org/10.1007/s00792-009-0231-9

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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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T1 - Structural determinants of the high thermal stability of SsoPox from the hyperthermophilic archaeon Sulfolobus solfataricus

AU - Del Vecchio, Pompea

AU - Elias, Mikael

AU - Merone, Luigia

AU - Graziano, Giuseppe

AU - Dupuy, Jérôme

AU - Mandrich, Luigi

AU - Carullo, Paola

AU - Fournier, Bertrand

AU - Rochu, Daniel

AU - Rossi, Mosè

AU - Masson, Patrick

AU - Chabriere, Eric

AU - Manco, Giuseppe

N1 - Funding Information: 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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N2 - 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.

AB - 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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KW - Quaternary structure organization

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

Abstract

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