Ca2+/calmodulin-dependent protein kinase II (CaMKII)

Abstract

The Ca2+/calmodulin (CaM)-dependent protein kinase II (CaMKII) is a major regulator of synaptic plasticity. CaMKIIfunction depends on complex regulation of its activity and localization by Ca2+/CaM and several auto-phosphorylation reactions. Auto-phosphorylation at T286 makes the kinase “autonomous” (partially active even without Ca2+/CaM), while auto-phosphorylation at T305/306 prevents subsequent Ca2+/CaM-binding. These processes also regulate synaptic localization and binding to the NMDA-receptor subunit GluN2B (formerly known as NR2B). Here we discuss studying CaMKII by inhibition (including by the novel class of CN inhibitors) and by mutagenesis. We describe purification of CaMKII, activity assays, directing and probing auto-phosphorylation, and investigating CaMKII protein-protein binding in vitro and within cells (with the GluN2B interaction as example).