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Domain structures of the PDK1 and PKCα constructs used in this study. Also shown are the N- or C-terminal affinity tags employed for protein purification and the ybbr-labeling tag used to attach Alexa Fluor 555 to PDK1 via Sfp enzymatic labeling. PKCα was unlabeled in the present study and was used as a dark binding partner. The AGC segment of PKCα contains the unstructured, tethered PIF motif that binds to the PIF pocket of the PDK1 kinase domain, as shown in Fig. 1.
