title: 2',3'-dideoxynucleoside triphosphates (ddNTP) and di-2',3'-dideoxynucleoside tetraphosphates (ddNp4ddN) behave differently to the corresponding NTP and Np4N counterparts as substrates of firefly luciferase, dinucleoside tetraphosphatase and phosphodiesterases.
abstract: 2',3'-Dideoxynucleosides (ddN) and their derivatives are currently used as antiretroviral compounds. Their active agents are the corresponding 2',3'-dideoxynucleoside triphosphates (ddNTPs) generated inside the cell by host kinases. Dinucleoside tetraphosphates (Np4Ns) are molecules of interest in metabolic regulation; their synthesis in vitro can be catalyzed by firefly luciferase. The relative synthesis of diadenosine 5',5'''-P1,P4-tetraphosphate or adenosine(5')tetraphospho(5')adenosine (Ap4A) from ATP is about 100-fold faster than that of di-2',3'-dideoxyadenosine 5',5'''-P1,P4-tetraphosphate or 2',3'-dideoxyadenosine (5')tetraphospho (5')-2',3'-dideoxyadenosine (ddAp4ddA) from ddATP. In the presence of ATPgammaS and ddATP the yield of adenosine(5')tetraphospo(5')-2',3'-dideoxyadenosine (Ap4ddA) was similar to that attained for Ap4A in the presence of ATP. The findings of this work indicate that the presence of a 3'-hydroxyl group is essential for the formation of the luciferase-luciferin-AMP complex, and explains the very low yield of ddAp4ddA in the presence of luciferase, luciferin and ddATP. The absence of 3'-hydroxyl groups in ddAp4ddA greatly hindered their hydrolysis by snake venom phosphodiesterase, asymmetrical dinucleoside tetraphosphatase and by a purified membrane preparation from rat liver. The possibility of using di-2',3'-dideoxynucleoside tetraphosphate (ddNp4ddN) or nucleoside(5')tetraphospho(5')-2',3'-dideoxynucleoside (Np4ddN) as a source of the active retroviral agent ddNTP, for example in HIV infection, is outlined.
authors.affiliation: Departamento de Bioquímica, Facultad de Medicina, Universidad Autónoma de Madrid, Spain.
authors.forename: M A
authors.initials: MA
authors.lastname: Sillero
authors.forename: O
authors.initials: O
authors.lastname: Madrid
authors.forename: E
authors.initials: E
authors.lastname: Zaera
authors.forename: A
authors.initials: A
authors.lastname: Sillero
mesh_terms: D017766:Acid Anhydride Hydrolases / Q000378:metabolism*; D000818:Animals; D000998:Antiviral Agents / Q000138:chemical synthesis*; D002462:Cell Membrane / Q000378:metabolism; D003854:Deoxyribonucleotides / Q000378:metabolism; D015224:Dideoxynucleosides / Q000378:metabolism; D015226:Dinucleoside Phosphates / Q000096:biosynthesis / Q000378:metabolism; D008099:Liver / Q000378:metabolism; D008156:Luciferases; D010727:Phosphoric Diester Hydrolases / Q000378:metabolism*; D051381:Rats
chemical_list: D000998:Antiviral Agents; D003854:Deoxyribonucleotides; D015224:Dideoxynucleosides; D015226:Dinucleoside Phosphates; C020733:diadenosine tetraphosphate; D008156:Luciferases; D010727:Phosphoric Diester Hydrolases; D017766:Acid Anhydride Hydrolases; C020023:bis(5'-nucleosyl)tetraphosphatase (asymmetrical)