[NMR paper] Direct Observation of Ca(2+) -Induced Calmodulin Conformational Transitions in Intact Xenopus laevis Oocytes by (19) F NMR Spectroscopy.

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Direct Observation of Ca(2+) -Induced Calmodulin Conformational Transitions in Intact Xenopus laevis Oocytes by (19) F NMR Spectroscopy.

Related Articles Direct Observation of Ca(2+) -Induced Calmodulin Conformational Transitions in Intact Xenopus laevis Oocytes by (19) F NMR Spectroscopy.

Angew Chem Int Ed Engl. 2015 Mar 5;

Authors: Ye Y, Liu X, Xu G, Liu M, Li C

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.


PMID: 25753548 [PubMed - as supplied by publisher]



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