Residual dipolar couplings (RDCs) provide valuable NMR parameters that can be used for structural calculation and verification. Measuring RDCs requires aligning macromolecules using one of various types of alignment media. Of different alignment media options, stretched or compressed polyacrylamide gels are advantageous due to their chemical stability. However, polyacrylamide interacts with proteins and significantly broadens NMR resonances. In this study, we found that the amide-containing compounds asparagine, glutamine and propionamide improve spectral quality of proteins in polyacrylamide gel without significantly reducing the magnitude of RDC values. Moreover, we showed that propionamide is an attractive additive that increases protein solubility without interfering with protein stability, ligand binding or NMR pulse width, suggesting its potential applications for our NMR methods.
[NMR paper] Sensitivity-enhanced Four-dimensional Amide-amide Correlation NMR Experiments for Sequential Assignment of Proline-rich Disordered Proteins.
Sensitivity-enhanced Four-dimensional Amide-amide Correlation NMR Experiments for Sequential Assignment of Proline-rich Disordered Proteins.
Sensitivity-enhanced Four-dimensional Amide-amide Correlation NMR Experiments for Sequential Assignment of Proline-rich Disordered Proteins.
J Am Chem Soc. 2018 Feb 28;:
Authors: Wong LE, Maier J, Wienands J, Becker S, Griesinger C
Abstract
Proline is prevalent in intrinsically disordered proteins (IDPs). NMR assignment of proline-rich IDPs is a challenge due to low dispersion of chemical...
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03-01-2018 09:20 PM
[NMR paper] A Unique and Simple Approach to Improve Sensitivity in (15)N-NMR Relaxation Measurements for NH?? Groups: Application to a Protein-DNA Complex.
A Unique and Simple Approach to Improve Sensitivity in (15)N-NMR Relaxation Measurements for NH?? Groups: Application to a Protein-DNA Complex.
Related Articles A Unique and Simple Approach to Improve Sensitivity in (15)N-NMR Relaxation Measurements for NH?? Groups: Application to a Protein-DNA Complex.
Molecules. 2017 Aug 15;22(8):
Authors: Nguyen D, Lokesh GLR, Volk DE, Iwahara J
Abstract
NMR spectroscopy is a powerful tool for research on protein dynamics. In the past decade, there has been significant progress in the...
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08-17-2017 01:01 PM
[NMR paper] Cross-Correlated Relaxation of Dipolar Coupling and Chemical-Shift Anisotropy in Magic-Angle Spinning R1? NMR Measurements: Application to Protein Backbone Dynamics Measurements.
Cross-Correlated Relaxation of Dipolar Coupling and Chemical-Shift Anisotropy in Magic-Angle Spinning R1? NMR Measurements: Application to Protein Backbone Dynamics Measurements.
Cross-Correlated Relaxation of Dipolar Coupling and Chemical-Shift Anisotropy in Magic-Angle Spinning R1? NMR Measurements: Application to Protein Backbone Dynamics Measurements.
J Phys Chem B. 2016 Aug 8;
Authors: Kurauskas V, Weber E, Hessel A, Ayala I, Marion D, Schanda P
Abstract
Transverse relaxation rate measurements in MAS solid-state NMR...
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08-09-2016 02:42 PM
Sequential backbone assignment based on dipolar amide-to-amide correlation experiments
Sequential backbone assignment based on dipolar amide-to-amide correlation experiments
Abstract
Proton detection in solid-state NMR has seen a tremendous increase in popularity in the last years. New experimental techniques allow to exploit protons as an additional source of information on structure, dynamics, and protein interactions with their surroundings. In addition, sensitivity is mostly improved and ambiguity in assignment experiments reduced. We show here that, in the solid state, sequential amide-to-amide correlations turn out to be an...
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05-15-2015 07:52 AM
[NMR paper] Characterization of polyacrylamide-stabilized Pfl phage liquid crystals for protein N
Characterization of polyacrylamide-stabilized Pfl phage liquid crystals for protein NMR spectroscopy.
Related Articles Characterization of polyacrylamide-stabilized Pfl phage liquid crystals for protein NMR spectroscopy.
J Biomol NMR. 2002 Jan;22(1):83-7
Authors: Trempe JF, Morin FG, Xia Z, Marchessault RH, Gehring K
A new polymer-stabilized nematic liquid crystal has been characterized for the measurement of biomolecular residual dipolar couplings. Filamentous Pf1 phage were embedded in a polyacrylamide matrix that fixes the orientation of...
Amide additives improve RDC measurements in polyacrylamide
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