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Chapitre D'ouvrage Année : 2020

Multiple Site-Specific Phosphorylation of IDPs Monitored by NMR

Résumé

In line with their high accessibility, disordered proteins are exquisite targets of kinases. Eukaryotic organisms use the so-called intrinsically disordered proteins (IDPs) or intrinsically disordered regions of proteins (IDRs) as molecular switches carrying intracellular information tuned by reversible phosphorylation schemes. Solvent-exposed serines and threonines are abundant in IDPs, and, consistently, kinases often modify disordered regions of proteins at multiple sites. In this context, nuclear magnetic resonance (NMR) spectroscopy provides quantitative, residue-specific information that permits mapping of phosphosites and monitoring of their individual kinetics. Hence, NMR monitoring emerges as an in vitro approach, complementary to mass-spectrometry or immuno-blotting, to characterize IDP phosphorylation comprehensively. Here, we describe in detail generic protocols for carrying out NMR monitoring of IDP phosphorylation, and we provide a number of practical insights that improve handiness and reproducibility of this method.
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Dates et versions

hal-02907472 , version 1 (07-12-2020)

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Manon Julien, Chafiaa Bouguechtouli, Ania Alik, Rania Ghouil, Sophie Zinn-Justin, et al.. Multiple Site-Specific Phosphorylation of IDPs Monitored by NMR. Intrinsically Disordered Proteins, 2141 (26), pp.793--817, 2020, ⟨10.1007/978-1-0716-0524-0_41⟩. ⟨hal-02907472⟩
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