Triple resonance 15N NMR relaxation experiments for studies of intrinsically disordered proteins
Result description
Description of protein dynamics is known to be essential in understanding their function. Studies based on a well established 15N NMR relaxation methodology have been applied to a large number of systems. However, the low dispersion of 1H chemical shifts very often observed within intrinsically disordered proteins complicates utilization of standard 2D HN correlated spectra because a limited number of amino acids can be characterized. Here we present a suite of triple resonance HNCO-type NMR experiments for measurements of five 15N relaxation parameters (R1, R2, NOE, cross-correlated relaxation rates gamma x and gamma z) in doubly 13C,15N-labeled proteins. We show that the third spectral dimension combined with non-uniform sampling provides relaxation rates for almost all residues of a protein with extremely poor chemical shift dispersion, the C terminal domain of delta-subunit of RNA polymerase from Bacillus subtilis. Comparison with data obtained using a sample labeled by 15N only showed that the presence of 13C has a negligible effect on gamma x, gamma z, and on the cross-relaxation rate (calculated from NOE and R1), and that these relaxation rates can be used to calculate accurate spectral density values. Partially 13C-labeled sample was used to test if the observed increase of 15NR1 in the presence of 13C corresponds to the 15N-13C dipole-dipole interactions in the 13C,15N-labeled sample.
Keywords
nuclear magnetic resonancerelaxationnon-uniform samplingintrinsically disordered proteins
The result's identifiers
Result code in IS VaVaI
Alternative codes found
RIV/68081715:_____/17:00483356 RIV/61388963:_____/17:00483356
Result on the web
DOI - Digital Object Identifier
Alternative languages
Result language
angličtina
Original language name
Triple resonance 15N NMR relaxation experiments for studies of intrinsically disordered proteins
Original language description
Description of protein dynamics is known to be essential in understanding their function. Studies based on a well established 15N NMR relaxation methodology have been applied to a large number of systems. However, the low dispersion of 1H chemical shifts very often observed within intrinsically disordered proteins complicates utilization of standard 2D HN correlated spectra because a limited number of amino acids can be characterized. Here we present a suite of triple resonance HNCO-type NMR experiments for measurements of five 15N relaxation parameters (R1, R2, NOE, cross-correlated relaxation rates gamma x and gamma z) in doubly 13C,15N-labeled proteins. We show that the third spectral dimension combined with non-uniform sampling provides relaxation rates for almost all residues of a protein with extremely poor chemical shift dispersion, the C terminal domain of delta-subunit of RNA polymerase from Bacillus subtilis. Comparison with data obtained using a sample labeled by 15N only showed that the presence of 13C has a negligible effect on gamma x, gamma z, and on the cross-relaxation rate (calculated from NOE and R1), and that these relaxation rates can be used to calculate accurate spectral density values. Partially 13C-labeled sample was used to test if the observed increase of 15NR1 in the presence of 13C corresponds to the 15N-13C dipole-dipole interactions in the 13C,15N-labeled sample.
Czech name
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Czech description
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Classification
Type
Jimp - Article in a specialist periodical, which is included in the Web of Science database
CEP classification
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OECD FORD branch
10606 - Microbiology
Result continuities
Project
Result was created during the realization of more than one project. More information in the Projects tab.
Continuities
P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)
Others
Publication year
2017
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Data specific for result type
Name of the periodical
Journal of Biomolecular NMR
ISSN
0925-2738
e-ISSN
—
Volume of the periodical
69
Issue of the periodical within the volume
3
Country of publishing house
NL - THE KINGDOM OF THE NETHERLANDS
Number of pages
14
Pages from-to
133-146
UT code for WoS article
000416856600003
EID of the result in the Scopus database
2-s2.0-85032189568
Basic information
Result type
Jimp - Article in a specialist periodical, which is included in the Web of Science database
OECD FORD
Microbiology
Year of implementation
2017