title: Structural rearrangement of CaMKIIalpha catalytic domains encodes activation.
abstract: At its fundamental level, human memory is thought to occur at individual synaptic contact sites and manifest as persistent changes in synaptic efficacy. In digital electronics, the fundamental structure for implementing memory is the flip-flop switch, a circuit that can be triggered to flip between two stable states. Recently, crystals of Ca(2+)/calmodulin-dependent protein kinase IIalpha (CaMKIIalpha) catalytic domains, the enzymatic portion of a dodecameric holoenzyme involved in memory, were found to form dimers [Rosenberg OS, Deindl S, Sung RJ, Nairn AC, Kuriyan J (2005) Structure of the autoinhibited kinase domain of CaMKII and SAXS analysis of the holoenzyme. Cell 123:849-860]. Although the formation of dimers in the intact holoenzyme has not been established, several features of the crystal structure suggest that dimers could act as a synaptic switch. ATP-binding sites were occluded, and the T286 autophosphorylation site responsible for persistent kinase activation was buried. These features would act to stabilize an autoinhibited "paired"-enzyme state. Ca(2+)-calmodulin binding was postulated to trigger the formation of an active state with unpaired catalytic domains. This conformation would allow ATP access and expose T286, autophosphorylation of which would act to maintain the "unpaired" conformation. We used fluorescence anisotropy and FRET imaging of Venus-tagged CaMKIIalpha to test the hypothesis that neuronal CaMKIIalpha can flip between two stable conformations in living cells. Our data support the existence of catalytic domain pairs, and glutamate receptor activation in neurons triggered an increase in anisotropy consistent with a structural transition from a paired to unpaired conformation.
authors.affiliation: National Institute on Alcohol Abuse and Alcoholism and Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, 5625 Fishers Lane, Rockville, MD 20892, USA.
authors.forename: Christopher
authors.initials: C
authors.lastname: Thaler
authors.forename: Srinagesh V
authors.initials: SV
authors.lastname: Koushik
authors.forename: Henry L
authors.initials: HL
authors.lastname: Puhl
authors.forename: Paul S
authors.initials: PS
authors.lastname: Blank
authors.forename: Steven S
authors.initials: SS
authors.lastname: Vogel