Triple resonance 15N NMR relaxation experiments for studies of intrinsically disordered proteins

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61388971%3A_____%2F17%3A00483356" target="_blank" >RIV/61388971:_____/17:00483356 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/68081715:_____/17:00483356 RIV/61388963:_____/17:00483356

Výsledek na webu

<a href="http://dx.doi.org/10.1007/s10858-017-0138-1" target="_blank" >http://dx.doi.org/10.1007/s10858-017-0138-1</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1007/s10858-017-0138-1" target="_blank" >10.1007/s10858-017-0138-1</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Triple resonance 15N NMR relaxation experiments for studies of intrinsically disordered proteins

Popis výsledku v původním jazyce

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.

Název v anglickém jazyce

Triple resonance 15N NMR relaxation experiments for studies of intrinsically disordered proteins

Popis výsledku anglicky

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.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

10606 - Microbiology

Projekt

Výsledek vznikl pri realizaci vícero projektů. Více informací v záložce Projekty.

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Rok uplatnění

2017

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Journal of Biomolecular NMR

ISSN

0925-2738

e-ISSN

—

Svazek periodika

69

Číslo periodika v rámci svazku

3

Stát vydavatele periodika

NL - Nizozemsko

Počet stran výsledku

14

Strana od-do

133-146

Kód UT WoS článku

000416856600003

EID výsledku v databázi Scopus

2-s2.0-85032189568