title: Conformational changes underlying calcium/calmodulin-dependent protein kinase II activation.
abstract: Calcium/calmodulin-dependent protein kinase II (CaMKII) interprets information conveyed by the amplitude and frequency of calcium transients by a controlled transition from an autoinhibited basal intermediate to an autonomously active phosphorylated intermediate (De Koninck and Schulman, 1998). We used spin labelling and electron paramagnetic resonance spectroscopy to elucidate the structural and dynamic bases of autoinhibition and activation of the kinase domain of CaMKII. In contrast to existing models, we find that autoinhibition involves a conformeric equilibrium of the regulatory domain, modulating substrate and nucleotide access. Binding of calmodulin to the regulatory domain induces conformational changes that release the catalytic cleft, activating the kinase and exposing an otherwise inaccessible phosphorylation site, threonine 286. Autophosphorylation at Thr286 further disrupts the interactions between the catalytic and regulatory domains, enhancing the interaction with calmodulin, but maintains the regulatory domain in a dynamic unstructured conformation following dissociation of calmodulin, sustaining activation. These findings support a mechanistic model of the CaMKII holoenzyme grounded in a dynamic understanding of autoregulation that is consistent with a wealth of biochemical and functional data.
authors.affiliation: Department of Molecular Physiology and Biophysics, Vanderbilt University School of Medicine, Nashville, TN 37232, USA.
authors.forename: Laurel
authors.initials: L
authors.lastname: Hoffman
authors.forename: Richard A
authors.initials: RA
authors.lastname: Stein
authors.forename: Roger J
authors.initials: RJ
authors.lastname: Colbran
authors.forename: Hassane S
authors.initials: HS
authors.lastname: Mchaourab
mesh_terms: D029742:Caenorhabditis elegans Proteins / Q000737:chemistry* / Q000378:metabolism*; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2 / Q000737:chemistry* / Q000378:metabolism*; D002147:Calmodulin / Q000378:metabolism*; D004578:Electron Spin Resonance Spectroscopy; D008954:Models, Biological; D008958:Models, Molecular; D010766:Phosphorylation; D011487:Protein Conformation
chemical_list: D029742:Caenorhabditis elegans Proteins; D002147:Calmodulin; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2