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

0483356 - ÚOCHB 2018 RIV NL eng J - Článek v odborném periodiku
Srb, Pavel - Nováček, J. - Kadeřávek, P. - Rabatinová, Alžběta - Krásný, Libor - Žídková, Jitka - Bobálová, Janette - Sklenář, V. - Žídek, L.
Triple resonance 15N NMR relaxation experiments for studies of intrinsically disordered proteins.
Journal of Biomolecular NMR. Roč. 69, č. 3 (2017), s. 133-146. ISSN 0925-2738. E-ISSN 1573-5001
Grant CEP: GA ČR GA13-16842S; GA MŠMT(CZ) LO1304
Institucionální podpora: RVO:61388963 ; RVO:61388971 ; RVO:68081715
Klíčová slova: nuclear magnetic resonance * relaxation * non-uniform sampling * intrinsically disordered proteins
Obor OECD: Analytical chemistry; Microbiology (MBU-M); Analytical chemistry (UIACH-O)
Impakt faktor: 2.534, rok: 2017

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.
Trvalý link: http://hdl.handle.net/11104/0278711