Identification, partial purification and inhibition by guanine analogues of a novel enzymic activity which phosphorylates guanosine to GMP in the protozoan parasite Eimeria tenella.
From oocysts of the protozoan parasite Eimeria tenella, responsible for avian coccidiosis, we have partially purified and characterized a novel enzymic activity which specifically phosphorylates guanosine to GMP. The enzyme is able to use several phosphate donors, in the order: acetyl phosphate (Ac-P) > ATP > UTP > CTP > phosphoribosyl pyrophosphate (PRPP) > dUTP > or = dATP. The low specificity of this enzyme for the phosphate donor suggested that it be named guanosine phosphotransferase (GPTase). This enzyme is biochemically distinct from the previously described adenosine kinase (AK) and hypoxanthine/xanthine/guanine phosphoribosyltransferase (HXGPRTase), and may enable the parasite to synthesize guanine nucleotides under conditions of imbalance between adenine and guanine nucleotides. Because of its possible role in the purine salvage pathways, we have studied the effect of several guanine and guanosine analogues, recently synthesized in our laboratory, on the activity of GPTase in vitro. GPTase is specifically inhibited in the micromolar range by several substituted N2-phenylguanine bases. These results indicate that, as previously found for AK and HXGPRTase, GPTase could be a potential target for antiparasitic chemotherapy.
Maga
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Istituto di Genetica Biochimica ed Evoluzionistica, CNR, Pavia, Italy.
Spadari
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Wright
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GE
Focher
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D000248:Adenosine Kinase / Q000037:antagonists & inhibitors; D000818:Animals; D000981:Antiprotozoal Agents / Q000494:pharmacology; D001345:Autoradiography; D002852:Chromatography, Ion Exchange; D016786:Eimeria tenella / Q000201:enzymology* / Q000378:metabolism; D004588:Electrophoresis, Cellulose Acetate; D004791:Enzyme Inhibitors; D006147:Guanine / Q000031:analogs & derivatives; D006151:Guanosine / Q000378:metabolism*; D006157:Guanosine Monophosphate / Q000378:metabolism*; D010766:Phosphorylation; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism*
D000981:Antiprotozoal Agents; D004791:Enzyme Inhibitors; D006151:Guanosine; D006147:Guanine; D006157:Guanosine Monophosphate; D017853:Phosphotransferases (Alcohol Group Acceptor); C086290:guanosine phosphotransferase; D000248:Adenosine Kinase