Unraveling the structural and functional differences between purine nucleoside phosphorylase and 5'-deoxy-5'-methylthioadenosine phosphorylase from the archaeon Pyrococcus furiosus. (Articolo in rivista)

Type

• Prodotto della ricerca (Classe)
• Articolo in rivista (Classe)

Label

• Unraveling the structural and functional differences between purine nucleoside phosphorylase and 5'-deoxy-5'-methylthioadenosine phosphorylase from the archaeon Pyrococcus furiosus. (Articolo in rivista) (literal)

Anno

• 2011-01-01T00:00:00+01:00 (literal)

Alternative label

• Cacciapuoti G, Marabotti A, Fuccio F, Porcelli M. (2011)
  Unraveling the structural and functional differences between purine nucleoside phosphorylase and 5'-deoxy-5'-methylthioadenosine phosphorylase from the archaeon Pyrococcus furiosus.
  in Biochemical and biophysical research communications (Print)
  (literal)

Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#autori

• Cacciapuoti G, Marabotti A, Fuccio F, Porcelli M. (literal)

Pagina inizio

• 1358 (literal)

Pagina fine

• 1366 (literal)

Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#numeroVolume

• 1814 (literal)

Rivista

• Biochemical and biophysical research communications (Rivista)

Note

• ISI Web of Science (WOS) (literal)

Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#affiliazioni

• Giovanna Cacciapuoti (Dip. Biochim. Biofis. Seconda Università di Napoli), Anna Marabotti (ITB-CNR & ISA-CNR), Francesca Fuccio (Dip. Biochim. Biofis. Seconda Università di Napoli), Marina Porcelli (Dip. Biochim. Biofis. Seconda Università di Napoli) (literal)

Titolo

• Unraveling the structural and functional differences between purine nucleoside phosphorylase and 5'-deoxy-5'-methylthioadenosine phosphorylase from the archaeon Pyrococcus furiosus. (literal)

Abstract

• Purine nucleoside metabolism in the archaeon Pyrococcus furiosus is catalyzed by purine nucleoside phosphorylase (PfPNP) and 5?-deoxy-5?-methylthioadenosine phosphorylase (PfMTAP). These enzymes, characterized by 50% amino acid sequence identity, show non-common features of thermophilicity and thermostability and are stabilized by intramolecular disulfide bonds. PfPNP is highly specific for 6-oxopurine nucleosides while PfMTAP is characterized by a broad substrate specificity with 6-aminopurine nucleosides as preferred substrates. Amino acid sequence comparison clearly shows that the hypothetical active sites of PfPNP and PfMTAP are almost identical and that, in analogy with human 5?-deoxy-5?-methylthioadenosine phosphorylase and human purine nucleoside phosphorylase, residue changes at level of the same crucial positions could be responsible for the switch of substrate specificity. To validate this hypothesis we changed the putative active site of PfPNP by site-directed mutagenesis. Substrate specificity and catalytic efficiency of PfPNP mutants were then analyzed by kinetic studies and compared with the wild-type enzyme. We carried out the molecular modeling of PfPNP and PfMTAP to obtain a picture of the overall enzyme structure and to identify structural features as well as interactions playing critical roles in thermostability. Finally, we utilized the structural models of mutant enzyme-substrate complex to rationalize the functional effects of the mutations. (literal)

Prodotto di

• Institute of food sciences (ISA) (Istituto)
• Bioinformatica per lo studio di patologie umane, intolleranze alimentari e sicurezza alimentare (INT.P02.006.001) (Modulo)

Autore CNR

• ANNA MARABOTTI (Unità di personale esterno)
• ANNA MARABOTTI (Unità di personale interno)

Incoming links:

Prodotto

• Institute of food sciences (ISA) (Istituto)
• Bioinformatica per lo studio di patologie umane, intolleranze alimentari e sicurezza alimentare (INT.P02.006.001) (Modulo)

Autore CNR di

• ANNA MARABOTTI (Unità di personale interno)
• ANNA MARABOTTI (Unità di personale esterno)

Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#rivistaDi

• Biochemical and biophysical research communications (Rivista)