An improved ultrafast 2D NMR experiment: Towards atom-resolved real-time studies of protein kinetics at multi-Hz rates |
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Authors: | Maayan Gal Thomas Kern Paul Schanda Lucio Frydman Bernhard Brutscher |
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Institution: | (1) Department of Chemical Physics, Weizmann Institute of Science, Rehovot, 76100, Israel;(2) Institut de Biologie Structurale, Jean-Pierre Ebel C.N.R.S.-C.E.A.-UJF 41, rue Jules Horowitz, Grenoble Cedex, 38027, France;(3) Laboratorium fuer Physikalische Chemie ETH Hoenggerberg, Zurich, CH, 8093, Switzerland |
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Abstract: | Multidimensional NMR spectroscopy is a well-established technique for the characterization of structure and fast-time-scale
dynamics of highly populated ground states of biological macromolecules. The investigation of short-lived excited states that
are important for molecular folding, misfolding and function, however, remains a challenge for modern biomolecular NMR techniques.
Off-equilibrium real-time kinetic NMR methods allow direct observation of conformational or chemical changes by following
peak positions and intensities in a series of spectra recorded during a kinetic event. Because standard multidimensional NMR
methods required to yield sufficient atom-resolution are intrinsically time-consuming, many interesting phenomena are excluded
from real-time NMR analysis. Recently, spatially encoded ultrafast 2D NMR techniques have been proposed that allow one to
acquire a 2D NMR experiment within a single transient. In addition, when combined with the SOFAST technique, such ultrafast
experiments can be repeated at high rates. One of the problems detected for such ultrafast protein NMR experiments is related
to the heteronuclear decoupling during detection with interferences between the pulses and the oscillatory magnetic field
gradients arising in this scheme. Here we present a method for improved ultrafast data acquisition yielding higher signal
to noise and sharper lines in single-scan 2D NMR spectra. In combination with a fast-mixing device, the recording of 1H–15N correlation spectra with repetition rates of up to a few Hertz becomes feasible, enabling real-time studies of protein kinetics
occurring on time scales down to a few seconds. |
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Keywords: | Spatially encoded NMR Ultrafast acquisition Multidimensional NMR Real-time NMR Hydrogen exchange Single-transition spin state |
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