Work

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

Year: 2017

Type: article

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 13C 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.

Source: Angewandte Chemie International Edition

DOI: 10.1002/anie.201700464

Authors: Mukundan Ragavan, Luigi I. Iconaru, Cheon‐Gil Park, Richard W. Kriwacki, Christian Hilty

Institutions: University of Florida Health, University of Florida, Texas A&M University, St. Jude Children's Research Hospital, University of Tennessee Health Science Center

Language: English

Topic: Advanced NMR Techniques and Applications

Subfield: Spectroscopy

Field: Chemistry

Domain: Physical Sciences

Open Access status: green

Funders: National Science Foundation, Welch Foundation, National Cancer Institute

Awards: G1020944575, CAREER: Biochemical Reaction Mechanisms by Real-Time, Hyperpolarization Enhanced Nuclear Magnetic Resonance, Characterization of Solvent-Macromolecule Interactions using Dissolution Dynamic Nuclear Polarization, Structural Determinants in Cell Growth Control by p21 and p27, Cancer Center Support Grant (CCSG)