FIGURE 2.

Sld2 and CDK-phosphorylated Sld2 bind to Mcm2–7 with similar affinity. A, 2 pmol of GST-Mcm2–7 (GST-Mcm2–7 has a GST tag at the N terminus of Mcm2), or GST was used to pull down various concentrations of radiolabeled Sld2 or CDK-phosphorylated Sld2. Bound protein was analyzed by SDS-PAGE and quantified with phosphorimaging. For detailed methods, see “Experimental Procedures.” B, results similar to the gels shown in A were averaged and plotted. C, experiments similar to A were conducted with 5 mm ATP, ATPγS, ADP, or no nucleotide. D, 2 pmol of GST-Sld2 or CDK-phosphorylated GST-Sld2 was used to pull down various concentrations of radiolabeled Mcm2–7 (radiolabeled Mcm2–7 has a radiolabeled PKA tag at the N terminus of Mcm3). E, the results were quantified, averaged, and plotted. F, experiments similar to D were conducted with 5 mm ATP, ATPγS, ADP, or no nucleotide.