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|Type:||Artigo de periódico|
|Title:||Characterisation Of A Gain-of-function Mutant Of Poly(adp-ribose) Polymerase|
De Murcia G.
De Murcia J.M.
|Abstract:||In order to examine the stucture-function relationship of the poly (ADP-ribose) polymerase (PARP) catalytic domain, potential active-site residues in the catalytic domain have previously been described. Here, we have used mutagenesis with hydroxylamine to generate a random library of PARP mutants. The identification, overproduction in insect cells, purification and characterization of a gain-of-function mutant (L713F) is described. We show that the k(cat) of this mutant is increased over nine times compared to the wild-type enzyme; the K(m) for NAD+ is unchanged. The size and the branching structure of the ADP-ribose polymers are similar in both the wild-type and the mutant enzyme. This mutation may have an allosteric effect on the catalytic site and could be useful in analyzing the consequences of poly ADP-ribose overproduction in vivo on cell survival following DNA damage.|
|Appears in Collections:||Unicamp - Artigos e Outros Documentos|
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