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Figure 5. Biochemical assays exclude the possibility of direct contact between ZD7288 and the CL-CNBD fragment and the interference with cAMP binding. (A) ZD7288 has no effect on the excitation (top) and emission (bottom) spectra of 8-NBD-cAMP in complex with purified HCN channel C-terminal protein. The effect of Mg2+ was tested separately, and no difference was found between 0 and 6 mM Mg2+. (B) ITC results showing that ZD7288 does not interfere with normal cAMP binding. (top) Raw trace showing the rate of heat exchange versus the time of cAMP injection (with 300 µM ZD7288). (bottom) Binding curves of cAMP to mHCN2 protein in the absence (black) and presence (red) of ZD7288. Image published in: Wu S et al. (2012) © 2012 Wu et al. Creative Commons Attribution-NonCommercial-ShareAlike license Larger Image Printer Friendly View

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