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Biochemical Analysis of AKAP-Anchored PKA Signaling Complexes.

Authors :
Byrne DP
Omar MH
Kennedy EJ
Eyers PA
Scott JD
Source :
Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2022; Vol. 2483, pp. 297-317.
Publication Year :
2022

Abstract

Generation of the prototypic second messenger cAMP instigates numerous signaling events. A major intracellular target of cAMP is Protein kinase A (PKA), a Ser/Thr protein kinase. Where and when this enzyme is activated inside the cell has profound implications on the functional impact of PKA. It is now well established that PKA signaling is focused locally into subcellular signaling "islands" or "signalosomes." The A-Kinase Anchoring Proteins (AKAPs) play a critical role in this process by dictating spatial and temporal aspects of PKA action. Genetically encoded biosensors, small molecule and peptide-based disruptors of PKA signaling are valuable tools for rigorous investigation of local PKA action at the biochemical level. This chapter focuses on approaches to evaluate PKA signaling islands, including a simple assay for monitoring the interaction of an AKAP with a tunable PKA holoenzyme. The latter approach evaluates the composition of PKA holoenzymes, in which regulatory subunits and catalytic subunits can be visualized in the presence of test compounds and small-molecule inhibitors.<br /> (© 2022. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

Details

Language :
English
ISSN :
1940-6029
Volume :
2483
Database :
MEDLINE
Journal :
Methods in molecular biology (Clifton, N.J.)
Publication Type :
Academic Journal
Accession number :
35286684
Full Text :
https://doi.org/10.1007/978-1-0716-2245-2_19