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cAMP Signaling pp 167-180 | Cite as

Structure-Based Bacteriophage Screening for AKAP-Selective PKA Regulatory Subunit Variants

  • Ryan Walker-Gray
  • Matthew G. GoldEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1294)

Abstract

cAMP-dependent protein kinase (PKA) is tethered at different subcellular locations by A-kinase anchoring proteins (AKAPs). AKAPs present amphipathic helices that bind to the docking and dimerization (D/D) domain of PKA regulatory subunits. Peptide disruptors derived from AKAP anchoring helices are powerful tools for determining whether PKA anchoring is important in different biological processes. Focusing on the reciprocal side of the AKAP-PKA interface can enable development of tools for determining the roles of individual AKAPs. Accordingly, here we describe a bacteriophage screening procedure for identifying variants of PKA regulatory subunit D/D domains that bind selectively to individual AKAPs. This procedure can be adapted for engineering specificity into other shared protein interfaces.

Keywords

PKA AKAP cAMP Phage display Protein engineering Directed evolution 

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  1. 1.Department of Neuroscience, Physiology and PharmacologyUniversity College LondonLondonUK

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