Angewandte Chemie · 2015 · 32 citations · 24 references
Life SciencesBiochemistryCell BiologyCell SignalingSignal TransductionStructural BiologyCalcium SignalingCalcium-binding ProteinsConformational DynamicsProtein DynamicsProtein StructureCalcium-calmodulin SignalingCalcium-dependent Conformational ChangeConformational Transition StatesParamagnetic Nmr SpectroscopyProtein Conformational DynamicsProtein Conformational SwitchingStructural ConstraintsTransition States In Conformational Changes
Abstract The Ca 2+ ‐mediated conformational transition of the protein calmodulin (CaM) is essential to a variety of signal transduction pathways. Whether the transition in living cells is similar to that observed in buffer is not known. Here, we report the direct observation by 19 F NMR spectroscopy of the transition of the Ca 2+ ‐free and ‐bound forms in Xenopus laevis oocytes at different Ca 2+ levels. We find that the Ca 2+ ‐bound CaM population increased greatly upon binding the target protein myosin light‐chain kinase (MLCK) at the same Ca 2+ level. Paramagnetic NMR spectroscopy was also exploited for the first time to obtain long‐range structural constraints in cells. Our study shows that 19 F NMR spectroscopy can be used to obtain long‐range structural constraints in living eukaryotic cells and paves the way for quantification of protein binding constants.
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NMR Structure Determination of Protein−Ligand Complexes by Lanthanide Labeling
Guido Pintacuda, Michael John, Xun‐Cheng Su et al. · Accounts of Chemical Research · 2007 · 272 citations
Impact of macromolecular crowding on DNA replication
Barak Akabayov, Sabine R. Akabayov, Seung‐Joo Lee et al. · Nature Communications · 2013 · 177 citations · Full text