Characterization of poly(ADP-ribose)polymerase from Crithidia fasciculata: enzyme inhibition by beta-lapachone.
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.
Villamil
S F
SF
Bioenergetics Research Centre (CONICET) and School of Medicine, University of Buenos Aires, Paraguay 2155, 1121-, Buenos Aires, Argentina.
Podestá
D
D
Molina Portela
M D
MD
Stoppani
A
A
D000818:Animals; D000981:Antiprotozoal Agents / Q000494:pharmacology*; D016831:Crithidia fasciculata / Q000201:enzymology*; D004591:Electrophoresis, Polyacrylamide Gel; D006861:Hydrogen Peroxide / Q000494:pharmacology; D007501:Iron / Q000494:pharmacology; D009285:Naphthoquinones / Q000494:pharmacology*; D000067856:Poly(ADP-ribose) Polymerase Inhibitors; D011065:Poly(ADP-ribose) Polymerases / Q000302:isolation & purification* / Q000378:metabolism*
D000981:Antiprotozoal Agents; C045076:Fenton's reagent; D009285:Naphthoquinones; D000067856:Poly(ADP-ribose) Polymerase Inhibitors; C014638:beta-lapachone; D006861:Hydrogen Peroxide; D007501:Iron; D011065:Poly(ADP-ribose) Polymerases