A-kinase Anchoring Proteins (AKAPs) are key orchestrators that spatiotemporally regulate protein kinase A (PKA) activity by scaffolding pertinent intracellular proteins to form signaling complexes. Although PKA and AKAPs are involved in a variety of cellular and physiological functions, their exact roles in these functions and corresponding regulation mechanism are not well understood. In this thesis, several peptides mimicking the A-Kinase Binding (AKB) helix of AKAPs were chemically stabilized using all-hydrocarbon stapling, in order to target the docking/dimerization domain of PKA-R in an isoform-selective manner. The peptides are cell permeable in diverse human cell lines, highly isoform-selective for PKA-RI or RII, and can effectively inhibit interactions between AKAPs and PKA-R in intact cells. Therefore, these peptides can be applied as valuable reagents in cell-based experiments to selectively disrupt AKAP-localized PKA-RI or -RII anchoring and aid in the study of compartmentalized type I or type II PKA signaling in cells.