Lobe-specific Functions of Ca2+·Calmodulin in αCa2+·Calmodulin-dependent Protein Kinase II Activation

摘要

N-Methyl-d-aspartic acid receptor-dependent long term potentiation (LTP), a model of memory formation, requires Ca²⁺·calmodulin-dependent protein kinase II (αCaMKII) activity and Thr²⁸⁶ autophosphorylation via both global and local Ca²⁺ signaling, but the mechanisms of signal transduction are not understood. We tested the hypothesis that the Ca²⁺-binding activator protein calmodulin (CaM) is the primary decoder of Ca²⁺ signals, thereby determining the output, e.g. LTP. Thus, we investigated the function of CaM mutants, deficient in Ca²⁺ binding at sites 1 and 2 of the N-terminal lobe or sites 3 and 4 of the C-terminal CaM lobe, in the activation of αCaMKII. Occupancy of CaM Ca²⁺ binding sites 1, 3, and 4 is necessary and sufficient for full activation. Moreover, the N- and C-terminal CaM lobes have distinct functions. Ca²⁺ binding to N lobe Ca²⁺ binding site 1 increases the turnover rate of the enzyme 5-fold, whereas the C lobe plays a dual role; it is required for full activity, but in addition, via Ca²⁺ binding site 3, it stabilizes ATP binding to αCaMKII 4-fold. Thr²⁸⁶ autophosphorylation is also dependent on Ca²⁺ binding sites on both the N and the C lobes of CaM. As the CaM C lobe sites are populated by low amplitude/low frequency (global) Ca²⁺ signals, but occupancy of N lobe site 1 and thus activation of αCaMKII requires high amplitude/high frequency (local) Ca²⁺ signals, lobe-specific sensing of Ca²⁺-signaling patterns by CaM is proposed to explain the requirement for both global and local Ca²⁺ signaling in the induction of LTP via αCaMKII.