A simple electrostatic switch important in the activation of type I protein kinase A by cyclic AMP.
Cyclic AMP activates protein kinase A by binding to an inhibitory regulatory (R) subunit and releasing inhibition of the catalytic (C) subunit. Even though crystal structures of regulatory and catalytic subunits have been solved, the precise molecular mechanism by which cyclic AMP activates the kinase remains unknown. The dynamic properties of the cAMP binding domain in the absence of cAMP or C-subunit are also unknown. Here we report molecular-dynamics simulations and mutational studies of the RIalpha R-subunit that identify the C-helix as a highly dynamic switch which relays cAMP binding to the helical C-subunit binding regions. Furthermore, we identify an important salt bridge which links cAMP binding directly to the C-helix that is necessary for normal activation. Additional mutations show that a hydrophobic "hinge" region is not as critical for the cross-talk in PKA as it is in the homologous EPAC protein, illustrating how cAMP can control diverse functions using the evolutionarily conserved cAMP-binding domains.
Vigil
Dominico
D
Department of Chemistry and Biochemistry, University of California, San Diego, San Diego, CA 92037, USA.
Lin
Jung-Hsin
JH
Sotriffer
Christoph A
CA
Pennypacker
Juniper K
JK
McCammon
J Andrew
JA
Taylor
Susan S
SS
D000818:Animals; D003198:Computer Simulation; D000242:Cyclic AMP / Q000737:chemistry / Q000378:metabolism*; D017868:Cyclic AMP-Dependent Protein Kinases / Q000737:chemistry / Q000378:metabolism*; D006358:Hot Temperature; D008956:Models, Chemical; D008958:Models, Molecular; D011489:Protein Denaturation; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D055672:Static Electricity
D000242:Cyclic AMP; D017868:Cyclic AMP-Dependent Protein Kinases