Measuring Solvent Hydrogen Exchange Rates by Multifrequency
Excitation 15N CEST: Application to Protein Phase Separation
Version 3 2018-09-25, 18:20
Version 2 2018-09-21, 18:04
Version 1 2018-09-19, 16:19
Posted on 2018-09-25 - 18:20
Solvent exchange rates provide important
information about the
structural and dynamical properties of biomolecules. A large number
of NMR experiments have been developed to measure such rates in proteins,
the great majority of which quantify the buildup of signals from backbone
amides after initial perturbation of water magnetization. Here we
present a different approach that circumvents the main limitations
that result from these classical hydrogen exchange NMR experiments.
Building on recent developments that enable rapid recording of chemical
exchange saturation transfer (CEST) pseudo-3D data sets, we describe
a 15N-based CEST scheme for measurement of solvent exchange
in proteins that exploits the one-bond 15N deuterium isotope
shift. The utility of the approach is verified with an application
to a 236 residue intrinsically disordered protein domain under conditions
where it phase separates and a second application involving a mutated
form of the domain that does not phase separate, establishing very
similar hydrogen exchange rates for both samples. The methodology
is well suited for studies of hydrogen exchange in any 15N-labeled biomolecule. A discussion of the merits of the CEST experiment
in relation to the popular CLEANEX-PM scheme is presented.
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Yuwen, Tairan; Bah, Alaji; Brady, Jacob P.; Ferrage, Fabien; Bouvignies, Guillaume; Kay, Lewis E. (2018). Measuring Solvent Hydrogen Exchange Rates by Multifrequency
Excitation 15N CEST: Application to Protein Phase Separation. ACS Publications. Collection. https://doi.org/10.1021/acs.jpcb.8b06820
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AUTHORS (6)
TY
Tairan Yuwen
AB
Alaji Bah
JB
Jacob P. Brady
FF
Fabien Ferrage
GB
Guillaume Bouvignies
LK
Lewis E. Kay
KEYWORDS
NMR experiments236 residueone-bond 15 N deuterium isotope shiftProtein Phase Separation Solvent exchange ratesCEST experimentMultifrequency Excitation 15 N CESTSolvent Hydrogen Exchange Ratesbackbone amideshydrogen exchange NMR experiments-3D data setshydrogen exchange ratesCLEANEX-PM schemehydrogen exchange15 N-based CEST schemewater magnetizationprotein domain15 N-labeled biomoleculechemical exchange saturation transfer