cAMP activation of PKA defines an ancient signaling mechanism

Rahul Das, Veronica Esposito, Mona Abu-Abed, Ganesh S. Anand, Susan S. Taylor, Giuseppe Melacini

Research output: Contribution to journalArticlepeer-review

105 Scopus citations

Abstract

cAMP and the cAMP binding domain (CBD) constitute a ubiquitous regulatory switch that translates an extracellular signal into a biological response. The CBD contains α- and β-subdomains with cAMP binding to a phosphate binding cassette (PBC) in the β-sandwich. The major receptors for cAMP in mammalian cells are the regulatory subunits (R-subunits) of PKA where cAMP and the catalytic subunit compete for the same CBD. The R-subunits inhibit kinase activity, whereas cAMP releases that inhibition. Here, we use NMR to map at residue resolution the cAMP-dependent interaction network of the CBD-A domain of isoform Iα of the R-subunit of PKA. Based on H/D, H/H, and Nz exchange data, we propose a molecular model for the allosteric regulation of PKA by cAMP. According to our model, cAMP binding causes long-range perturbations that propagate well beyond the immediate surroundings of the PBC and involve two key relay sites located at the C terminus of β2 (1163) and N terminus of β3 (D170). The I163 site functions as one of the key triggers of global unfolding, whereas the D170 locus acts as an electrostatic switch that mediates the communication between the PBC and the B-helix. Removal of cAMP not only disrupts the cap for the B′ helix within the PBC, but also breaks the circuitry of cooperative interactions stemming from the PBC, thereby uncoupling the α- and β-subdomains. The proposed model defines a signaling mechanism, conserved in every genome, where allosteric binding of a small ligand disrupts a large protein-protein interface.

Original languageEnglish (US)
Pages (from-to)93-98
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume104
Issue number1
DOIs
StatePublished - Jan 2 2007

All Science Journal Classification (ASJC) codes

  • General

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