engleski

Steady-state kinetic analysis of E. coli purine nucleoside phosphorylase active site mutants

In order to get close insight into very complex mechanism of Escherichia coli purine nucleoside phosphorylase (PNP) kinetic measurements of a wild type enzyme and its Arg24Ala mutant towards phosphate as substrate were performed and discussed. The key role of Arg24 during phosphorolysis was studied at different pH and at constant nucleoside (7-methylguanosine and adenosine) concentration. The comparative analysis of the catalytic sites of wild type enzyme and Arg24Ala mutant based on kinetic data will be presented. E. coli PNP, in contrast to human homologue, has a broad substrate specificity which makes this enzyme a good candidate in gene therapy against solid tumours. Therefore, understanding of its catalytic mechanism is of utmost importance.

purine nucleoside phosphorylase; active site mutants; steady-state kinetics

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