Structural insights into the calcium-mediated allosteric transition in the C-terminal domain of calmodulin from NMR measurements.
Related Articles Structural insights into the calcium-mediated allosteric transition in the C-terminal domain of calmodulin from NMR measurements.
Biochemistry. 2015 Nov 30;
Authors: Kukic P, Lundström P, Camilloni C, Evenäs J, Akke M, Vendruscolo M
Abstract
Calmodulin is a two-domain signalling protein that becomes activated upon binding cooperatively two pairs of calcium ions, leading to large-scale conformational changes that expose its binding site. Despite significant advances in understanding the structural biology of calmodulin function, the mechanistic details of the conformational transition between closed and open states have remained unclear. To investigate this transition, we used a combination of molecular dynamics simulations and NMR experiments on the Ca2+-saturated E140Q C-terminal domain variant. Using chemical shift restraints in replica-averaged metadynamics simulations, we obtained a high-resolution structural ensemble consisting of two conformational states, and validated such ensemble against three independent experimental datasets, namely inter-proton NOEs, 15N order parameters, and chemical shift differences between the exchanging states. Through a detailed analysis of this structural ensemble and of the corresponding statistical weights, we characterised a calcium-mediated conformational transition whereby the coordination of Ca2+ by just one oxygen of the bidentate ligand E140 triggers a concerted movement of the two EF-hands that exposes the target-binding site. This analysis provides atomistic insights into a possible Ca2+-mediated activation mechanism of calmodulin not achievable from static structures alone or from ensemble NMR measurements of the transition between conformations.
PMID: 26618792 [PubMed - as supplied by publisher]
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