TY - JOUR T1 - Enzymatic Activity of CaMKII is not Required for its Interaction with the Glutamate Receptor Subunit GluN2B JF - Molecular Pharmacology JO - Mol Pharmacol DO - 10.1124/mol.113.089045 SP - mol.113.089045 AU - Kelsey Barcomb AU - Steven J Coultrap AU - K. Ulrich Bayer Y1 - 2013/01/01 UR - http://molpharm.aspetjournals.org/content/early/2013/10/15/mol.113.089045.abstract N2 - Binding of the Ca2+/calmodulin(CaM)-dependent protein kinase II (CaMKII) to the NMDA-type glutamate receptor (NMDAR) subunit GluN2B is an important control mechanism for the regulation of synaptic strength. CaMKII binding to GluN2B and CaMKII translocation to synapses is induced by an initial Ca2+/CaM-stimulus, which also activates the kinase. Indeed, several mechanistically different CaMKII inhibitors (tatCN21 and KN93) and inactivating mutations (K42M, A302R, and T305/T306D) impair this interaction, suggesting that it requires CaMKII enzymatic activity. However, this study shows that two general kinase inhibitors H7 and staurosporine (Sta), which inhibit CaMKII activity by yet another mechanism, did not interfere with GluN2B binding in vitro or within cells. In contrast to a previous report, we found that Sta, like H7, inhibited CaMKII in an ATP-competitive manner. Nucleotide binding significantly enhances CaMKII/GluN2B binding in vitro, but the nucleotide-competition by H7 or Sta did not prevent this effect and instead even mimicked it. H7 (700 μM) and Sta (2 μM) efficiently blocked enzymatic activity of CaMKII both in vitro and within cells. However, neither H7 nor Sta prevented Ca2+-induced translocation of CaMKII to GluN2B in heterologous cells or to synapses in hippocampal neurons. Thus, activity of CaMKII (or of any other kinase inhibited by H7 or Sta) is not required for stimulation-induced GluN2B-binding and synaptic translocation of CaMKII, despite previous indication to the contrary. This shows that results with inhibitors and inhibiting mutants can be caused by structural effects independent from catalytic activity, and that detailed understanding of the mechanisms is required for their interpretation. ER -