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Real‐Time Analysis of Folding upon Binding of a Disordered Protein by Using Dissolution DNP NMR Spectroscopy
Author(s) -
Ragavan Mukundan,
Iconaru Luigi I.,
Park CheonGil,
Kriwacki Richard W.,
Hilty Christian
Publication year - 2017
Publication title -
angewandte chemie international edition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.831
H-Index - 550
eISSN - 1521-3773
pISSN - 1433-7851
DOI - 10.1002/anie.201700464
Subject(s) - nuclear magnetic resonance spectroscopy , dissolution , folding (dsp implementation) , spectroscopy , protein folding , chemistry , crystallography , nuclear magnetic resonance , stereochemistry , physics , biochemistry , engineering , electrical engineering , quantum mechanics
The kinase inhibitory domain of the cell cycle regulatory protein p27 Kip1 (p27) was nuclear spin hyperpolarized using dissolution dynamic nuclear polarization (D‐DNP). While intrinsically disordered in isolation, p27 adopts secondary structural motifs, including an α‐helical structure, upon binding to cyclin‐dependent kinase 2 (Cdk2)/cyclin A. The sensitivity gains obtained with hyperpolarization enable the real‐time observation of 13 C NMR signals during p27 folding upon binding to Cdk2/cyclin A on a time scale of several seconds. Time‐dependent intensity changes are dependent on the extent of folding and binding, as manifested in differential spin relaxation. The analysis of signal decay rates suggests the existence of a partially folded p27 intermediate during the timescale of the D‐DNP NMR experiment.