Biochemical characterization and homology modeling of a purine-specific ribonucleoside hydrolase from the archaeon Sulfolobus solfataricus: Insights into mechanisms of protein stabilization

Marina Porcelli, Iolanda Peluso, Anna Marabotti, Angelo Facchiano, Giovanna Cacciapuoti

Research output: Contribution to journalArticlepeer-review

Abstract

We report the biochemical and structural characterization of the purine-specific ribonucleoside hydrolase from the archaeon Sulfolobus solfataricus (SsIAG-NH). SsIAG-NH is a homodimer of 70 kDa specific for adenosine, guanosine and inosine. SsIAG-NH is highly thermophilic and is characterized by extreme thermodynamic stability (Tm, 107 °C), kinetic stability and remarkable resistance to guanidinium chloride-induced unfolding. A disulfide bond that, on the basis of SDS-PAGE is positioned intersubunits, plays an important role in thermal stability. SsIAG-NH shares 43% sequence identity with the homologous pyrimidine-specific nucleoside hydrolase from S. solfataricus (SsCU-NH). The comparative sequence alignment of SsIAG-NH, SsCU-NH, purine non-specific nucleoside hydrolase from Crithidia fasciculata and purine-specific nucleoside hydrolase from Trypanosoma vivax shows that, only few changes in the base pocket are responsible for different substrate specificity of two S. solfataricus enzymes. The structure of SsIAG-NH predicted by homology modeling allows us to infer the role of specific residues in substrate specificity and thermostability.

Original languageEnglish
Pages (from-to)55-65
Number of pages11
JournalArchives of Biochemistry and Biophysics
Volume483
Issue number1
DOIs
Publication statusPublished - Mar 1 2009

Keywords

  • Archaea
  • Disulfide bonds
  • Homology modeling
  • Nucleoside hydrolase
  • Nucleoside metabolism
  • Sulfolobus solfataricus
  • Thermostability

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

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