Effect of GMPPNP and GDP on tRNA ligation activity of PF0027 protein. The effect of GMPPNP on (A) GTP-dependent and (B) GTPγS-dependent tRNA ligation activities and the effect of (C) GDP on GTP-dependent tRNA ligation activity were examined. The reaction was carried out with 4 μg/mL purified PF0027 protein for 15 min at 37°C with the indicated concentration of GMPPNP, GTP, or GDP. Synthetic tRNA halves bearing 2′,3′-cyclic phosphate and 5′-OH termini were used for all the experiments. (Arrowheads) Indicate the positions of ligated product (tRNA). (Bottom) For competition experiments, the relative amounts (%) of ligation products are indicated. Similar results were obtained from at least two independent experiments.
