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Probing Invisible, Excited Protein States by Non‐Uniformly Sampled Pseudo‐4D CEST Spectroscopy
Author(s) -
Long Dong,
Delaglio Frank,
Sekhar Ashok,
Kay Lewis E.
Publication year - 2015
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.201504070
Subject(s) - limiting , spectroscopy , spectral line , excited state , chemistry , saturation (graph theory) , resolution (logic) , nuclear magnetic resonance spectroscopy , high resolution , nuclear magnetic resonance , analytical chemistry (journal) , computational physics , biological system , molecular physics , physics , computer science , atomic physics , mathematics , artificial intelligence , chromatography , mechanical engineering , remote sensing , quantum mechanics , astronomy , combinatorics , engineering , biology , geology
Chemical exchange saturation transfer (CEST) NMR spectroscopy is a powerful tool for studies of slow timescale protein dynamics. Typical experiments are based on recording a large number of 2D data sets and quantifying peak intensities in each of the resulting planes. A weakness of the method is that peaks must be resolved in 2D spectra, limiting applications to relatively small proteins. Resolution is significantly improved in 3D spectra but recording uniformly sampled data is time‐prohibitive. Here we describe non‐uniformly sampled HNCO‐based pseudo‐4D CEST that provides excellent resolution in reasonable measurement times. Data analysis is done through fitting in the time domain, without the need of reconstructing the frequency dimensions, exploiting previously measured accurate peak positions in reference spectra. The methodology is demonstrated on several protein systems, including a nascent form of superoxide dismutase that is implicated in neurodegenerative disease.