Real-Time Analysis of Folding upon Binding of a Disordered Protein by Using Dissolution DNP NMR Spectroscopy

Angewandte Chemie
Mukundan RagavanChristian Hilty

Abstract

The kinase inhibitory domain of the cell cycle regulatory protein p27Kip1 (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.

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Citations

Mar 31, 2018·Magnetic Resonance in Chemistry : MRC·Guannan Zhang, Christian Hilty
Jul 10, 2019·Physical Chemistry Chemical Physics : PCCP·Koki NishimuraNobuhiro Yanai
Jun 5, 2020·Chemical Communications : Chem Comm·Koki NishimuraNobuhiro Yanai
Jan 18, 2020·Proceedings of the National Academy of Sciences of the United States of America·Mihajlo NovakovicLucio Frydman
May 31, 2019·Proceedings of the National Academy of Sciences of the United States of America·Yusuke OkunoSilvia Cavagnero
Mar 5, 2021·Emerging Topics in Life Sciences·Hong ZhangSarah Perrett
Mar 14, 2021·Journal of Magnetic Resonance·Jihyun KimChristian Hilty

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