Unambiguous Tracking of Protein Phosphorylation by Fast High‐Resolution FOSY NMR**

Autor: Wolfgang Bermel, Eduard V. Bocharov, Vladislav Yu. Orekhov, Björn M. Burmann, Irena Matečko-Burmann, Panagiota S. Georgoulia, Dmitry M. Lesovoy, Tammo Diercks
Rok vydání: 2021
Předmět:
Zdroj: Angewandte Chemie (International Ed. in English)
ISSN: 1521-3757
0044-8249
Popis: Dysregulation of post‐translational modifications (PTMs) like phosphorylation is often involved in disease. NMR may elucidate exact loci and time courses of PTMs at atomic resolution and near‐physiological conditions but requires signal assignment to individual atoms. Conventional NMR methods for this base on tedious global signal assignment that may often fail, as for large intrinsically disordered proteins (IDPs). We present a sensitive, robust alternative to rapidly obtain only the local assignment near affected signals, based on FOcused SpectroscopY (FOSY) experiments using selective polarisation transfer (SPT). We prove its efficiency by identifying two phosphorylation sites of glycogen synthase kinase 3 beta (GSK3β) in human Tau40, an IDP of 441 residues, where the extreme spectral dispersion in FOSY revealed unprimed phosphorylation also of Ser409. FOSY may broadly benefit NMR studies of PTMs and other hotspots in IDPs, including sites involved in molecular interactions.
A new suite of 2D FOSY experiments focuses on one coupled nuclear spin system at a time, that is, a PTM site or a structural hotspot in general, with 3–5 known frequencies. With higher sensitivity and versatility than achievable by traditional experiments, FOSY solves the spectral dispersion problem and obtains only the relevant local NMR signal assignment in a few hours, as demonstrated for two phosphorylation sites in the 441‐residue IDP Tau.
Databáze: OpenAIRE
Externí odkaz:
Nepřihlášeným uživatelům se plný text nezobrazuje