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Probing the interaction of a quercetin bioconjugate with Bcl‐2 in living human cancer cells with in‐cell NMR spectroscopy
Author(s) -
Primikyri Alexandra,
Sayyad Nisar,
Quilici Giacomo,
Vrettos Eirinaios I.,
Lim Kyungeun,
Chi SeungWook,
Musco Giovanna,
Gerothanassis Ioannis P.,
Tzakos Andreas G.
Publication year - 2018
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1002/1873-3468.13250
Subject(s) - bioconjugation , intracellular , chemistry , cancer cell , nuclear magnetic resonance spectroscopy , cell , protein–protein interaction , biophysics , biochemistry , cancer , biology , stereochemistry , genetics
In‐cell NMR spectroscopy has emerged as a powerful technique for monitoring biomolecular interactions at an atomic level inside intact cells. However, current methodologies are inadequate at charting intracellular interactions of nonlabeled proteins and require their prior isotopic labeling. Herein, we describe for the first time the monitoring of the quercetin‐alanine bioconjugate interaction with the nonlabeled antiapoptotic protein Bcl‐2 inside living human cancer cells. STD and Tr‐ NOESY in‐cell NMR methodologies were successfully applied in the investigation of the binding, which was further validated in vitro . In‐cell NMR proved a very promising strategy for the real‐time probing of the interaction profile of potential drugs with their therapeutic targets in native cellular environments and could, thus, open a new avenue in drug discovery.