Bioinformatic design of A-kinase anchoring protein-in silico: a potent and selective peptide antagonist of type II protein kinase A anchoring.
Compartmentalization of the cAMP-dependent protein kinase (PKA) is coordinated through association with A-kinase anchoring proteins (AKAPs). A defining characteristic of most AKAPs is a 14- to 18-aa sequence that binds to the regulatory subunits (RI or RII) of the kinase. Cellular delivery of peptides to these regions disrupts PKA anchoring ... and has been used to delineate a physiological role for AKAPs in the facilitation of certain cAMP-responsive events. Here, we describe a bioinformatic approach that yields an RII-selective peptide, called AKAP-in silico (AKAP-IS), that binds RII with a K(d) of 0.4 nM and binds RI with a K(d) of 277 nM. AKAP-IS associates with the type II PKA holoenzyme inside cells and displaces the kinase from natural anchoring sites. Electrophysiological recordings indicate that perfusion of AKAP-IS evokes a more rapid and complete attenuation of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor currents than previously described anchoring inhibitor peptides. Thus, computer-based and peptide array screening approaches have generated a reagent that binds PKA with higher affinity than previously described AKAPs.
Mesh Terms:
Amino Acid Sequence, Base Sequence, Cell Line, Computational Biology, Consensus Sequence, Cyclic AMP-Dependent Protein Kinase Type II, Cyclic AMP-Dependent Protein Kinases, DNA, Complementary, Drug Design, Humans, Kinetics, Models, Molecular, Molecular Sequence Data, Protein Conformation, Protein Engineering, Proteins, Recombinant Proteins, Signal Transduction
Amino Acid Sequence, Base Sequence, Cell Line, Computational Biology, Consensus Sequence, Cyclic AMP-Dependent Protein Kinase Type II, Cyclic AMP-Dependent Protein Kinases, DNA, Complementary, Drug Design, Humans, Kinetics, Models, Molecular, Molecular Sequence Data, Protein Conformation, Protein Engineering, Proteins, Recombinant Proteins, Signal Transduction
Proc. Natl. Acad. Sci. U.S.A.
Date: Apr. 15, 2003
PubMed ID: 12672969
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