Fig. 5. Cps2D ATP-binding domain is required for direct interaction with SpAdK.
a SDS-PAGE gels showing purified recombinant GST-SpAdK, His6-Cps2D wild-type (WT), 3Y3E (Y215E/Y218E/Y221E), GK-AA (G48A/K49A), and GK-AA/3Y3E. b BLI sensorgrams of the interactions between SpAdK and Cps2D WT or 3Y3E in the presence or absence of ATP. For clarity, only sensorgrams at the maximal concentration of Cps2D protein (50 μM) are shown. c BLI sensorgrams of the interaction between SpAdK with Cps2D. His6-Cps2D was immobilized on Ni-NTA sensors, followed by the addition of varying concentrations of GST-SpAdK (2.5, 5, 10, 25, and 50 μM). RU response unit. d BLI-derived binding curves for the interactions of SpAdK with Cps2D WT, 3Y3E, GK-AA, and GK-AA/3Y3E mutants for steady-state binding analysis. The data are representative of two independent experiments