title: Uncoupling Catalytic and Binding Functions in the Cyclic AMP-Dependent Protein Kinase A.
abstract: The canonical function of kinases is to transfer a phosphoryl group to substrates, initiating a signaling cascade; while their non-canonical role is to bind other kinases or substrates, acting as scaffolds, competitors, and signal integrators. Here, we show how to uncouple kinases' dual function by tuning the binding cooperativity between nucleotide (or inhibitors) and substrate allosterically. We demonstrate this new concept for the C subunit of protein kinase A (PKA-C). Using thermocalorimetry and nuclear magnetic resonance, we found a linear correlation between the degree of cooperativity and the population of the closed state of PKA-C. The non-hydrolyzable ATP analog (ATPγC) does not follow this correlation, suggesting that changing the chemical groups around the phosphoester bond can uncouple kinases' dual function. Remarkably, this uncoupling was also found for two ATP-competitive inhibitors, H89 and balanol. Since the mechanism for allosteric cooperativity is not conserved in different kinases, these results may suggest new approaches for designing selective kinase inhibitors.
authors.affiliation: Department of Chemistry, University of Minnesota, Minneapolis, MN 55455, USA; Department of Biochemistry, Molecular Biology and Biophysics, University of Minnesota, Minneapolis, MN 55455, USA.
authors.forename: Jonggul
authors.initials: J
authors.lastname: Kim
authors.affiliation: Department of Chemistry, University of Minnesota, Minneapolis, MN 55455, USA.
authors.forename: Geoffrey
authors.initials: G
authors.lastname: Li
authors.affiliation: Department of Medicinal Chemistry, Institute for Therapeutics Discovery and Development, University of Minnesota, Minneapolis, MN 55455, USA.
authors.forename: Michael A
authors.initials: MA
authors.lastname: Walters
authors.affiliation: Department of Chemistry and Biochemistry, University of California at San Diego, La Jolla, CA 92093, USA.
authors.forename: Susan S
authors.initials: SS
authors.lastname: Taylor
authors.affiliation: Department of Chemistry, University of Minnesota, Minneapolis, MN 55455, USA; Department of Biochemistry, Molecular Biology and Biophysics, University of Minnesota, Minneapolis, MN 55455, USA. Electronic address: vegli001@umn.edu.
authors.forename: Gianluigi
authors.initials: G
authors.lastname: Veglia
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism*; D000818:Animals; D001381:Azepines / Q000494:pharmacology; D002151:Calorimetry; D020134:Catalytic Domain; D054751:Cyclic AMP-Dependent Protein Kinase Catalytic Subunits / Q000737:chemistry* / Q000378:metabolism*; D006801:Humans; D062385:Hydroxybenzoates / Q000494:pharmacology; D007546:Isoquinolines / Q000494:pharmacology; D008279:Magnetic Resonance Imaging; D008958:Models, Molecular; D009711:Nucleotides / Q000378:metabolism*; D011485:Protein Binding; D011487:Protein Conformation; D013449:Sulfonamides / Q000494:pharmacology
chemical_list: D001381:Azepines; D062385:Hydroxybenzoates; D007546:Isoquinolines; D009711:Nucleotides; D013449:Sulfonamides; C015405:ophiocordin; D000255:Adenosine Triphosphate; D054751:Cyclic AMP-Dependent Protein Kinase Catalytic Subunits; C063509:N-(2-(4-bromocinnamylamino)ethyl)-5-isoquinolinesulfonamide
keywords: canonical and non-canonical function; cooperativity; phospholamban; protein kinase A; pseudo-kinases
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