Abstract
Nucleoside hydrolases are metalloproteins that hydrolyze the N-glycosidic bond of β-ribonucleosides, forming the free purine/pyrimidine base and ribose. We report the stability of the two hyperthermophilic enzymes Sulfolobus solfataricus pyrimidine-specific nucleoside hydrolase (SsCU-NH) and Sulfolobus solfataricus purine-specific inosineadenosine- guanosine nucleoside hydrolase (SsIAG-NH) against the denaturing action of temperature and guanidine hydrochloride by means of circular dichroism and fluorescence spectroscopy. The guanidine hydrochloride-induced unfolding is reversible for both enzymes as demonstrated by the analysis of the refolding process by activity assays and fluorescence measurements. The evidence that the denaturation of SsIAG-NH carried out in the presence of reducing agents proved to be reversible indicates that the presence of disulfide bonds interferes with the refolding process of this enzyme. Both enzymes are highly thermostable and no thermal unfolding transition can be obtained up to 108°C. SsIAG-NH is thermally denatured under reducing conditions (Tm=93°C) demonstrating the contribution of disulfide bridges to enzyme thermostability.
Keywords: Circular dichroism, Conformational stability, disulfide bonds, hyperthermophilic enzymes, nucleoside hydrolases, Sulfolobus solfataricus, prokaryotes, SsCU-NH, pyrimidine, thermostability
Protein & Peptide Letters
Title: Thermal Unfolding of Nucleoside Hydrolases from the Hyperthermophilic Archaeon Sulfolobus solfataricus: Role of Disulfide Bonds
Volume: 19 Issue: 3
Author(s): Marina Porcelli, Ester De Leo, Pompea Del Vecchio, Francesca Fuccio and Giovanna Cacciapuoti
Affiliation:
Keywords: Circular dichroism, Conformational stability, disulfide bonds, hyperthermophilic enzymes, nucleoside hydrolases, Sulfolobus solfataricus, prokaryotes, SsCU-NH, pyrimidine, thermostability
Abstract: Nucleoside hydrolases are metalloproteins that hydrolyze the N-glycosidic bond of β-ribonucleosides, forming the free purine/pyrimidine base and ribose. We report the stability of the two hyperthermophilic enzymes Sulfolobus solfataricus pyrimidine-specific nucleoside hydrolase (SsCU-NH) and Sulfolobus solfataricus purine-specific inosineadenosine- guanosine nucleoside hydrolase (SsIAG-NH) against the denaturing action of temperature and guanidine hydrochloride by means of circular dichroism and fluorescence spectroscopy. The guanidine hydrochloride-induced unfolding is reversible for both enzymes as demonstrated by the analysis of the refolding process by activity assays and fluorescence measurements. The evidence that the denaturation of SsIAG-NH carried out in the presence of reducing agents proved to be reversible indicates that the presence of disulfide bonds interferes with the refolding process of this enzyme. Both enzymes are highly thermostable and no thermal unfolding transition can be obtained up to 108°C. SsIAG-NH is thermally denatured under reducing conditions (Tm=93°C) demonstrating the contribution of disulfide bridges to enzyme thermostability.
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Cite this article as:
Porcelli Marina, De Leo Ester, Del Vecchio Pompea, Fuccio Francesca and Cacciapuoti Giovanna, Thermal Unfolding of Nucleoside Hydrolases from the Hyperthermophilic Archaeon Sulfolobus solfataricus: Role of Disulfide Bonds, Protein & Peptide Letters 2012; 19 (3) . https://dx.doi.org/10.2174/092986612799363091
DOI https://dx.doi.org/10.2174/092986612799363091 |
Print ISSN 0929-8665 |
Publisher Name Bentham Science Publisher |
Online ISSN 1875-5305 |
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