Sensitivity-Enhanced 13C-NMR for Monitoring Multisite Phosphorylation at Physiological Temperature and pH.
Angew Chem Int Ed Engl. 2020 Mar 17;:
Authors: Theillet FX, Zinn-Justin S, Alik A, Bouguechtouli C, Julien M, Ghouil R, Bermel W
Abstract
Abundant phosphorylation events rule the activity of nuclear proteins involved in gene regulation and DNA repair. These occur mostly on disordered regions of proteins, which often contain multiple phosphosites. Comprehensive and quantitative monitoring of phosphorylation reactions is theoretically achievable at a residue-specific level using 1 H- 15 N NMR spectroscopy, but is often limited by low signal-to-noise at pH>7 and T>293K. We developed an improved 13 C?- 13 CO correlation NMR experiment that works equally at any pH or temperature, i.e. also at those where genuine kinase activities occur. This permits to obtain atomic-resolution information in physiological conditions down to 25 ?M. We exemplified the interest of this approach by monitoring phosphorylation reactions, in presence of purified kinases or in cell extracts, on a range of previously problematic targets, namely Mdm2, BRCA2 and Oct4.
PMID: 32181947 [PubMed - as supplied by publisher]
[ASAP] Structural Regulation of a Neurofilament-Inspired Intrinsically Disordered Protein Brush by Multisite Phosphorylation
Structural Regulation of a Neurofilament-Inspired Intrinsically Disordered Protein Brush by Multisite Phosphorylation
https://pubs.acs.org/appl/literatum/publisher/achs/journals/content/bichaw/0/bichaw.ahead-of-print/acs.biochem.8b00007/20180405/images/medium/bi-2018-00007d_0006.gif
Biochemistry
DOI: 10.1021/acs.biochem.8b00007
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Perdeuterated and 13C-enriched myo-inositol for DNP assisted monitoring of enzymatic phosphorylation by inositol-3-kinase
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Moure, M.J., et al., Perdeuterated and 13C-enriched myo-inositol for DNP assisted monitoring of enzymatic phosphorylation by inositol-3-kinase. Chemical Communications, 2017. 53(92): p. 12398-12401.
http://dx.doi.org/10.1039/C7CC07023C
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Abstract
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Sensitivity-Enhanced 13C-NMR for Monitoring Multisite Phosphorylation at Physiological Temperature and pH.
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