The C-terminal V5 domain of Protein Kinase C is intrinsically disordered, with propensity to associate with a membrane mimetic.
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The C-terminal V5 domain is one of the most variable domains in Protein Kinase C isoforms (PKCs). V5 confers isoform specificity on its parent enzyme through interactions with isoform-specific adaptor proteins and possibly through specific intra-molecular interactions with other PKC domains. The structural information about V5 domains in solution is sparse. The objective of this work was to determine the conformational preferences of the V5 domain from the isoform of PKC (V5) and evaluate its ability to associate with membrane mimetics. We show that V5 and its phosphorylation-mimicking variant, dmV5, are intrinsically disordered protein domains. Phosphorylation-mimicking mutations do not alter the overall conformation of the polypeptide backbone, as evidenced by the local nature of chemical shift perturbations and the secondary structure propensity scores. However, the population of the "cis-trans" conformer of the Thr(638)-Pro(639)-Pro(640) turn motif, which has been implicated in the down-regulation of PKC via peptidyl-prolyl isomerase Pin1, increases in dmV5, along with the conformational flexibility of the region between the turn and hydrophobic motifs. Both wild type and dmV5 associate with micelles made of a zwitterionic detergent, n-dodecylphosphocholine. Upon micelle binding, V5 acquires a higher propensity to form helical structures at the conserved "NFD" motif and the entire C-terminal third of the domain. The ability of V5 to partition into the hydrophobic micellar environment suggests that it may serve as a membrane anchor during the PKC maturation process.