Direct observation of Ca(2+) -induced calmodulin conformational transitions in intact Xenopus laevis oocytes by (19) F NMR spectroscopy

Angewandte Chemie
Yansheng YeConggang Li

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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Citations

Jul 30, 2016·Chemical Communications : Chem Comm·Bin-Bin PanXun-Cheng Su
May 6, 2019·Chemphyschem : a European Journal of Chemical Physics and Physical Chemistry·Arina DalaloyanDaniella Goldfarb
Dec 1, 2017·Analytical and Bioanalytical Chemistry·Yansheng YeConggang Li
Aug 31, 2019·Chemical Communications : Chem Comm·Wenwen ZhengConggang Li
May 22, 2019·International Journal of Molecular Sciences·Teppei IkeyaYutaka Ito
Oct 24, 2017·Organic & Biomolecular Chemistry·Brittany A RiggleIvan J Dmochowski
Dec 11, 2017·Journal of Biomolecular NMR·Jia-Liang ChenXun-Cheng Su
Mar 17, 2017·Annual Review of Analytical Chemistry·Conggang LiMaili Liu
Jan 14, 2021·Analytical Chemistry·Yunfei HuMaili Liu
Aug 30, 2017·Biochimica Et Biophysica Acta. General Subjects·Teppei IkeyaChristian Griesinger
Apr 11, 2021·Proceedings of the National Academy of Sciences of the United States of America·Shannon L SpeerConggang Li
May 20, 2019·Biochimica Et Biophysica Acta. General Subjects·Noritaka NishidaIchio Shimada
Apr 22, 2021·Current Opinion in Chemical Biology·Fabio Arnesano
May 3, 2019·The Journal of Physical Chemistry. B·Yansheng YeConggang Li

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