Related ArticlesNMR Relaxation Mechanisms for Backbone Carbonyl Carbons in a 13 C, 15 N-Labeled Protein
J Magn Reson. 1997 May;126(1):48-57
Authors: Allard P, Härd T
The predominant relaxation mechanisms for backbone carbonyl carbon (13 C') relaxation in a 13 C, 15 N-doubly enriched sample of the thermostable Sso7d protein have been investigated. Pulse sequences for measurements of longitudinal and transverse 13 C' relaxation rates were implemented, and these rates were measured at magnetic fields of 11.7 and 14.1 T. The field dependence in measured rates is small and consistent with a predominant contribution from chemical-shift anisotropy (CSA) to 13 C' relaxation. A pulse sequence for measurement of {1 H}-13 C' cross-relaxation rates (steady-state NOEs) was also developed. This experiment reveals a significant NOE between protons and all 13 C', indicating that dipolar interactions between these nuclei contribute to 13 C' relaxation. Experiments designed to suppress cross correlation between CSA relaxation and dipole-dipole (DD) relaxation due to neighboring 13 Calpha indicate that this effect is negligible. A more quantitative treatment is also presented, in which backbone dynamics parameters are fitted to average 13 C' relaxation rates using Lipari-Szabo expressions for the spectral density. This fit, which reproduces well expected backbone dynamics parameters for a folded protein, is used to estimate the relative contributions of various mechanisms to 13 C' relaxation. It is found that both longitudinal and transverse relaxation rates are dominated by CSA relaxation and contain significant contributions due to DD relaxation induced by nearby protons. Contributions from DD relaxation due to covalently bound 13 Calpha and 15 N are comparably small. The predominant effects of CSA and 1 H-13 C' DD interactions, for which physical and geometrical parameters are uncertain, complicate the use of 13 C' relaxation as a sequence-specific probe for protein backbone dynamics.
[NMR paper] Temperature dependence of protein backbone motion from carbonyl 13C and amide 15N NMR
Temperature dependence of protein backbone motion from carbonyl 13C and amide 15N NMR relaxation.
Related Articles Temperature dependence of protein backbone motion from carbonyl 13C and amide 15N NMR relaxation.
J Magn Reson. 2005 May;174(1):43-53
Authors: Chang SL, Tjandra N
The NMR spin-lattice relaxation rate (R1) and the rotating-frame spin-lattice relaxation rate (R1rho) of amide 15N and carbonyl 13C (13C') of the uniformly 13C- and 15N-labeled ubiquitin were measured at different temperatures and field strengths to investigate the...
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[NMR paper] Auto-induction medium for the production of [U-15N]- and [U-13C, U-15N]-labeled prote
Auto-induction medium for the production of - and -labeled proteins for NMR screening and structure determination.
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Protein Expr Purif. 2005 Apr;40(2):268-78
Authors: Tyler RC, Sreenath HK, Singh S, Aceti DJ, Bingman CA, Markley JL, Fox BG
Protocols have been developed and applied for the high-throughput production of - or -labeled proteins using the conditional methionine auxotroph Escherichia coli B834. The...
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[NMR paper] Suppression of diagonal peaks in TROSY-type 1H NMR NOESY spectra of 15N-labeled prote
Suppression of diagonal peaks in TROSY-type 1H NMR NOESY spectra of 15N-labeled proteins.
Related Articles Suppression of diagonal peaks in TROSY-type 1H NMR NOESY spectra of 15N-labeled proteins.
J Magn Reson. 1999 Oct;140(2):499-503
Authors: Meissner A, Sørensen OW
A novel method for suppression of diagonal peaks in the amide region of NOESY NMR spectra of 15N-labeled proteins is presented. The method is particularly useful for larger proteins at high magnetic fields where interference between dipolar and chemical shift anisotropy relaxation...
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11-18-2010 08:31 PM
[NMR paper] NMR Relaxation Mechanisms for Backbone Carbonyl Carbons in a 13 C, 15 N-Labeled Prote
NMR Relaxation Mechanisms for Backbone Carbonyl Carbons in a 13 C, 15 N-Labeled Protein
http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--linkinghub.elsevier.com-ihub-images-PubMedLink.gif Related Articles NMR Relaxation Mechanisms for Backbone Carbonyl Carbons in a 13 C, 15 N-Labeled Protein
J Magn Reson. 1997 May;126(1):48-57
Authors: Allard P, Härd T
The predominant relaxation mechanisms for backbone carbonyl carbon (13 C') relaxation in a 13 C, 15 N-doubly enriched sample of the thermostable Sso7d protein have been investigated....
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08-22-2010 03:03 PM
1H-13C Separated Local Field Spectroscopy of Uniformly 13C Labeled Peptides and Prote
1H-13C Separated Local Field Spectroscopy of Uniformly 13C Labeled Peptides and Proteins
Publication year: 2010
Source: Journal of Magnetic Resonance, In Press, Accepted Manuscript, Available online 1 July 2010</br>
Eugene C., Lin , Chin H., Wu , Yuan, Yang , Christopher V., Grant , Stanley J., Opella</br>
By incorporating homonuclear decoupling on both the 1H and 13C channels it is feasible to obtain high-resolution two-dimensional separated local field spectra of peptides and proteins that are 100% labeled with 13C. Dual-PISEMO (Polarization Inversion Spin Exchange Modulated...
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08-16-2010 03:50 AM
HA-detected experiments for the backbone assignment of intrinsically disordered prote
Abstract We propose a new alpha proton detection based approach for the sequential assignment of natively unfolded proteins. The proposed protocol superimposes on following features: HA-detection (1) enables assignment of natively unfolded proteins at any pH, i.e., it is not sensitive to rapid chemical exchange undergoing in natively unfolded proteins even at moderately high pH. (2) It allows straightforward assignment of proline-rich polypeptides without additional proline-customized experiments. (3) It offers more streamlined and less ambiguous assignment based on solely intraresidual...
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08-14-2010 04:19 AM
Comparison of fast backbone dynamics at amide nitrogen and carbonyl sites in dematin
Abstract We perform a detailed comparison of fast backbone dynamics probed at amide nitrogen versus carbonyl carbon sites for dematin headpiece C-terminal domain (DHP) and its S74E mutant (DHPS74E). Carbonyl dynamics is probed via auto-correlated longitudinal rates and transverse Câ?²/Câ?²-Cα CSA/dipolar and Câ?²/Câ?²â??N CSA/dipolar cross-correlated rates, while 15N data are taken from a previous study. Resulting values of effective order parameters and internal correlation times support the conclusion that Câ?² relaxation reports on a different subset of fast motions compared to those...
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Measurement of carbonyl chemical shifts of excited protein states by relaxation dispersion NMR spectroscopy: comparison between uniformly and selectively 13C labeled samples
Measurement of carbonyl chemical shifts of excited protein states by relaxation dispersion NMR spectroscopy: comparison between uniformly and selectively 13C labeled samples
Patrik Lundström, D. Flemming Hansen and Lewis E. Kay
Journal of Biomolecular NMR; 2008; 42(1); pp 35 - 47
Abstract: Carr–Purcell–Meiboom–Gill (CPMG) relaxation dispersion nuclear magnetic resonance (NMR) spectroscopy has emerged as a powerful method for quantifying chemical shifts of excited protein states. For many applications of the technique that involve the measurement of relaxation rates of carbon...