Data di Pubblicazione:
2004
Citazione:
Mutagenesis of Escherichia coli uridine phosphorylase by random pentapeptide insertions / I. Oliva, G. Zuffi, G. Orsini, G. Tonon, L. De Gioia, D. Ghisotti. - In: ENZYME AND MICROBIAL TECHNOLOGY. - ISSN 0141-0229. - 35:4(2004), pp. 309-314. [10.1093/jb/mvhO57]
Abstract:
Escherichia coli uridine phosphorylase (UP) is encoded by the udp gene and catalyzes the reversible phosphorolysis of uridine to uracil and ribose-1-phosphate. Only few key residues involved in UP catalysis, identified by site-directed mutagenesis and selective chemical modification studies, were reported. A recent paper reporting the crystal structure at 2.0 Angstrom resolution indicated that UP shares a high structural homology with E. coli purine nucleoside phosphorylase. This latter enzyme is better known and a number of residues in its active site have been identified. In this work, we used pentapeptide scanning mutagenesis to cause random insertions of a 5 amino acid cassette in the UP polypeptide chain. Several insertional mutants located in different regions of UP maintained or increased the enzymatic activity and provided new insights into protein structure-function relationships. Moreover, this mutagenesis approach appears to be useful for the rapid preparation of mutants that present altered enzymatic activities.
Tipologia IRIS:
01 - Articolo su periodico
Keywords:
uridine phosphorylase ; Escherichia coli ; mutagenesis ; transposon
Elenco autori:
I. Oliva, G. Zuffi, G. Orsini, G. Tonon, L. De Gioia, D. Ghisotti
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