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Characterization of poly(ADP-ribose)polymerase from Crithidia fasciculata: enzyme inhibition by beta-lapachone
S F Villamil1, D Podestá, M D Molina Portela
1Bioenergetics Research Centre (CONICET) and School of Medicine, University of Buenos Aires, Paraguay 2155, 1121-, Buenos Aires, Argentina.
Abstract:
Crithidia fasciculata poly(ADP-ribose)polymerase (PARP) has been isolated and partially purified. This is the first PARP isolated from trypanosomatids; it requires DNA and histone for activity, using NAD(+) as substrate. Thiol compounds specially dithiothreitol essentially contributed to PARP stability during purification and to PARP activity during assays. Nicotinamide, 3-aminobenzamide, theophylline, histamine, histidine, N-ethylmaleimide, p-chloromercuribenzoic acid, p-chloromercuriphenylsulfonic acid and o-iodosobenzoate inhibited PARP, thus confirming enzyme identity. PARP was also inhibited by the Fe(II)/H(2)O(2) Fenton system. beta-Lapachone inhibited PARP, apparently by direct interaction with the enzyme.
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