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Interaction of the retinal G-protein transducin with uracil nucleotides
1Institut für Pharmakologie, Freie Universität Berlin, Thielallee 69-73, Berlin, D-14195, Germany.
Abstract:
Little is known about the interaction of pyrimidine nucleotides with G-proteins. Here we report that under experimental conditions that exclude transphosphorylation reactions, nucleoside 5'-triphosphates inhibited transducin-catalyzed GTP hydrolysis in the order of potency guanosine 5'-[gamma-thio]triphosphate > GTP > guanosine 5'-[beta,gamma-imido]triphosphate > uridine 5'-[gamma-thio]triphosphate > UTP > CTP. Nucleoside 5'-diphosphates inhibited GTP hydrolysis in the order of potency GDP approximately guanosine 5'-[beta-thio]thiodiphosphate > uridine 5'-[beta-thio]diphosphate >> UDP (no effect). UTP inhibited GTP hydrolysis competitively, indicative for nucleotide binding to the same site. Uracil nucleotides had a distinct activity profile with respect to disruption of the transitory complex between photoexcited rhodopsin and nucleotide-free transducin. We conclude that (i) uracil nucleotides bind to transducin-alpha with lower affinity than the corresponding guanine nucleotides, (ii) phosphorothioate modification of uracil nucleotides increases their affinity for transducin, and (iii) uracil nucleotides induce conformational changes in G-proteins that are different from the conformational changes induced by guanine nucleotides.
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