title: PKA-type I selective constrained peptide disruptors of AKAP complexes.
abstract: A-Kinase Anchoring Proteins (AKAPs) coordinate complex signaling events by serving as spatiotemporal modulators of cAMP-dependent protein kinase activity in cells. Although AKAPs organize a plethora of diverse pathways, their cellular roles are often elusive due to the dynamic nature of these signaling complexes. AKAPs can interact with the type I or type II PKA holoenzymes by virtue of high-affinity interactions with the R-subunits. As a means to delineate AKAP-mediated PKA signaling in cells, we sought to develop isoform-selective disruptors of AKAP signaling. Here, we report the development of conformationally constrained peptides named RI-STapled Anchoring Disruptors (RI-STADs) that target the docking/dimerization domain of the type 1 regulatory subunit of PKA. These high-affinity peptides are isoform-selective for the RI isoforms, can outcompete binding by the classical AKAP disruptor Ht31, and can selectively displace RIα, but not RIIα, from binding the dual-specific AKAP149 complex. Importantly, these peptides are cell-permeable and disrupt Type I PKA-mediated phosphorylation events in the context of live cells. Hence, RI-STAD peptides are versatile cellular tools to selectively probe anchored type I PKA signaling events.
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.affiliation: †Department of Pharmaceutical and Biomedical Sciences, College of Pharmacy, University of Georgia, Athens, Georgia 30602, United States.
authors.forename: Tienhuei G
authors.initials: TG
authors.lastname: Ho
authors.affiliation: ‡Department of Biochemistry, University of Kassel, 34132 Kassel, Germany.
authors.forename: Eugen
authors.initials: E
authors.lastname: Franz
authors.affiliation: ‡Department of Biochemistry, University of Kassel, 34132 Kassel, Germany.
authors.forename: Jennifer S
authors.initials: JS
authors.lastname: Hermann
authors.affiliation: §Howard Hughes Medical Institute, Department of Pharmacology, University of Washington School of Medicine, Seattle, Washington 98195, United States.
authors.forename: F Donelson
authors.initials: FD
authors.lastname: Smith
authors.affiliation: §Howard Hughes Medical Institute, Department of Pharmacology, University of Washington School of Medicine, Seattle, Washington 98195, United States.
authors.forename: Heidi
authors.initials: H
authors.lastname: Hehnly
authors.affiliation: §Howard Hughes Medical Institute, Department of Pharmacology, University of Washington School of Medicine, Seattle, Washington 98195, United States.
authors.forename: Jessica L
authors.initials: JL
authors.lastname: Esseltine
authors.affiliation: †Department of Pharmaceutical and Biomedical Sciences, College of Pharmacy, University of Georgia, Athens, Georgia 30602, United States.
authors.forename: Laura E
authors.initials: LE
authors.lastname: Hanold
authors.affiliation: †Department of Pharmaceutical and Biomedical Sciences, College of Pharmacy, University of Georgia, Athens, Georgia 30602, United States.
authors.forename: Mandi M
authors.initials: MM
authors.lastname: Murph
authors.affiliation: ‡Department of Biochemistry, University of Kassel, 34132 Kassel, Germany.
authors.forename: Daniela
authors.initials: D
authors.lastname: Bertinetti
authors.affiliation: §Howard Hughes Medical Institute, Department of Pharmacology, University of Washington School of Medicine, Seattle, Washington 98195, United States.
authors.forename: John D
authors.initials: JD
authors.lastname: Scott
authors.affiliation: ‡Department of Biochemistry, University of Kassel, 34132 Kassel, Germany.
authors.forename: Friedrich W
authors.initials: FW
authors.lastname: Herberg
authors.affiliation: †Department of Pharmaceutical and Biomedical Sciences, College of Pharmacy, University of Georgia, Athens, Georgia 30602, United States.
authors.forename: Eileen J
authors.initials: EJ
authors.lastname: Kennedy
mesh_terms: D054758:A Kinase Anchor Proteins / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism; D000595:Amino Acid Sequence; D001665:Binding Sites / Q000187:drug effects; D045744:Cell Line, Tumor; D002463:Cell Membrane Permeability; D054743:Cyclic AMP-Dependent Protein Kinase Type I / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism; D054746:Cyclic AMP-Dependent Protein Kinase Type II / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry* / Q000494:pharmacology; D010766:Phosphorylation; D011485:Protein Binding / Q000187:drug effects; D011487:Protein Conformation; D047428:Protein Kinase Inhibitors / Q000737:chemistry* / Q000494:pharmacology; D021122:Protein Subunits / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism
chemical_list: D054758:A Kinase Anchor Proteins; C105137:AKAP1 protein, human; D010455:Peptides; D047428:Protein Kinase Inhibitors; D021122:Protein Subunits; D054743:Cyclic AMP-Dependent Protein Kinase Type I; D054746:Cyclic AMP-Dependent Protein Kinase Type II