Figure 4.

Relative g5p-binding affinity of BA-8 versus BA-8-C. (A) EMSA. A fixed amount of ³²P-labeled BA-8 (1 μM) was mixed with increasing concentrations of unlabeled BA-8-C (0–107 μM) in a buffer of 20 mM Tris–HCl, pH 7.4, 200 mM KCl and 1.5 mM MgCl₂. After adding 11.2 μM of g5p, the mixtures were incubated at 37°C for 15 min and subjected to a 2.5% agarose gel. Shown is a phosphor image of the dried gel. Lane 1, ³²P-labeled BA-8 only. (B) Quantitation of complex formation inhibited by BA-8-C. Percentages of g5p·BA-8 complexes retained in lanes 2–10 of (A) were quantitated and normalized to the sample without BA-8-C (lane 2).