TAILIEUCHUNG - Báo cáo khoa học: Pyrophosphate and tripolyphosphate affect firefly luciferase luminescence because they act as substrates and not as allosteric effectors

The activating and stabilizing effects of inorganic pyrophosphate, tripoly-phosphate and nucleoside triphosphates on firefly luciferase biolumines-cence were studied. The results obtained show that those effects are a consequence of the luciferase-catalyzed splitting of dehydroluciferyl-adenyl-ate, a powerful inhibitor formed as a side product in the course of the bio-luminescence reaction. | ễFEBS Journal Pyrophosphate and tripolyphosphate affect firefly luciferase luminescence because they act as substrates and not as allosteric effectors Rui Fontes1 Diogo Fernandes1 2 Filipe Peralta2 Hugo Fraga2 Ines Maio1 and Joaquim C. G. Esteves da Silva2 1 Servico de Bioquimica U38-FCT Faculdade de Medicina da Universidade do Porto Portugal 2 Departamento de Quimica Faculdade de Ciencias da Universidade do Porto Centro de Investigacao em Quimica UP Portugal Keywords dehydroluciferyl-adenylate dinucleoside polyphosphates firefly luciferase pyrophosphate tripolyphosphate Correspondence J. C. G. Esteves da Silva Departamento de Quimica Faculdade de Ciencias da Universidade do Porto R. Campo Alegre 687 4169 007 Porto Portugal Fax 351 220 402 659 Tel 351 220 402 569 E-mail jcsilva@ Present address Department of Cellular Biology Harvard MedicalSchool Boston MA USA Received 30 October 2007 revised 15 January 2008 accepted 21 January 2008 doi The activating and stabilizing effects of inorganic pyrophosphate tripolyphosphate and nucleoside triphosphates on firefly luciferase bioluminescence were studied. The results obtained show that those effects are a consequence of the luciferase-catalyzed splitting of dehydroluciferyl-adenyl-ate a powerful inhibitor formed as a side product in the course of the bioluminescence reaction. Inorganic pyrophosphate tripolyphosphate CTP and UTP antagonize the inhibitory effect of dehydroluciferyl-adenylate because they react with it giving rise to products that are at least less powerful inhibitors. Moreover we demonstrate that the antagonizing effects depended on the rate of the splitting reactions being higher in the cases of inorganic pyrophosphate and tripolyphosphate and lower in the cases of CTP and UTP. In the case of inorganic pyrophosphate the correlation between the rate of dehydroluciferyl-adenylate pyrophosphorolysis and the activating effect on bioluminescence only occurs for low .

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