title: Isoform-selective disruption of AKAP-localized PKA using hydrocarbon stapled peptides.
abstract: A-kinase anchoring proteins (AKAPs) play an important role in the spatial and temporal regulation of protein kinase A (PKA) by scaffolding critical intracellular signaling complexes. Here we report the design of conformationally constrained peptides that disrupt interactions between PKA and AKAPs in an isoform-selective manner. Peptides derived from the A Kinase Binding (AKB) domain of several AKAPs were chemically modified to contain an all-hydrocarbon staple and target the docking/dimerization domain of PKA-R, thereby occluding AKAP interactions. The peptides are cell-permeable against diverse human cell lines, are highly isoform-selective for PKA-RII, and can effectively inhibit interactions between AKAPs and PKA-RII in intact cells. These peptides can be applied as useful reagents in cell-based studies to selectively disrupt AKAP-localized PKA-RII activity and block AKAP signaling complexes. In summary, the novel hydrocarbon-stapled peptides developed in this study represent a new class of AKAP disruptors to study compartmentalized RII-regulated PKA signaling in cells.
authors.affiliation: Department of Pharmaceutical and Biomedical Sciences, College of Pharmacy, University of Georgia , Athens, Georgia 30602, United States.
authors.forename: Yuxiao
authors.initials: Y
authors.lastname: Wang
authors.forename: Tienhuei G
authors.initials: TG
authors.lastname: Ho
authors.forename: Daniela
authors.initials: D
authors.lastname: Bertinetti
authors.forename: Matthias
authors.initials: M
authors.lastname: Neddermann
authors.forename: Eugen
authors.initials: E
authors.lastname: Franz
authors.forename: Gary C H
authors.initials: GC
authors.lastname: Mo
authors.forename: Lewis P
authors.initials: LP
authors.lastname: Schendowich
authors.forename: Avinash
authors.initials: A
authors.lastname: Sukhu
authors.forename: Raybun C
authors.initials: RC
authors.lastname: Spelts
authors.forename: Jin
authors.initials: J
authors.lastname: Zhang
authors.forename: Friedrich W
authors.initials: FW
authors.lastname: Herberg
authors.forename: Eileen J
authors.initials: EJ
authors.lastname: Kennedy
mesh_terms: D054758:A Kinase Anchor Proteins / Q000737:chemistry / Q000378:metabolism*; D000595:Amino Acid Sequence; D045744:Cell Line, Tumor; D057846:Cell-Penetrating Peptides / Q000737:chemistry*; D017868:Cyclic AMP-Dependent Protein Kinases / Q000737:chemistry / Q000378:metabolism*; D005454:Fluorescence Polarization; D006801:Humans; D047468:Immunoprecipitation; D008856:Microscopy, Fluorescence; D008958:Models, Molecular; D008969:Molecular Sequence Data; D019151:Peptide Library; D011485:Protein Binding; D020033:Protein Isoforms; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D015398:Signal Transduction; D013379:Substrate Specificity
chemical_list: D054758:A Kinase Anchor Proteins; D057846:Cell-Penetrating Peptides; D019151:Peptide Library; D020033:Protein Isoforms; D011993:Recombinant Fusion Proteins; D017868:Cyclic AMP-Dependent Protein Kinases