Abstract We present new NMR methods to measure slow translational diffusion coefficients of biomolecules. Like the heteronuclear stimulated echo experiment (XSTE), these new methods rely on the storage of information about spatial localization during the diffusion delay as longitudinal polarization of nuclei with long T1 such as nitrogen-15. The new BEST-XSTE sequence combines features of Band-selective Excitation Short-Transient (BEST) and XSTE methods. By avoiding the saturation of all protons except those of amide groups, one can increase the sensitivity by 45% in small proteins. The new experiment which combines band-Selective Optimized Flip-Angle Short-Transient with XSTE (SOFAST-XSTE) offers an alternative when very short recovery delays are desired. A modification of the HSQC-edited version of the XSTE experiment offers enhanced sensitivity and access to higher resolution in the indirect dimension. These new methods have been applied to detect changes in diffusion coefficients due to dimerization or proteolysis of Engrailed 2, a partially disordered protein.
Content Type Journal Article
Pages 1-10
DOI 10.1007/s10858-011-9510-8
Authors
Rafal Augustyniak, Département de chimie, Ecole Normale Supérieure, 24 rue Lhomond, 75231 Paris Cedex 05, France
Fabien Ferrage, Département de chimie, Ecole Normale Supérieure, 24 rue Lhomond, 75231 Paris Cedex 05, France
Raphaël Paquin, Département de chimie, Ecole Normale Supérieure, 24 rue Lhomond, 75231 Paris Cedex 05, France
Olivier Lequin, Département de chimie, Ecole Normale Supérieure, 24 rue Lhomond, 75231 Paris Cedex 05, France
Geoffrey Bodenhausen, Département de chimie, Ecole Normale Supérieure, 24 rue Lhomond, 75231 Paris Cedex 05, France
Paramagnetic relaxation enhancement to improve sensitivity of fast NMR methods: application to intrinsically disordered proteins
Paramagnetic relaxation enhancement to improve sensitivity of fast NMR methods: application to intrinsically disordered proteins
Abstract We report enhanced sensitivity NMR measurements of intrinsically disordered proteins in the presence of paramagnetic relaxation enhancement (PRE) agents such as Ni2+-chelated DO2A. In proton-detected 1H-15N SOFAST-HMQC and carbon-detected (H-flip)13CO-15N experiments, faster longitudinal relaxation enables the usage of even shorter interscan delays. This results in higher NMR signal intensities per units of experimental time, without adverse line...
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Spin Diffusion Driven by R-Symmetry Sequences: Applications to Homonuclear Correlation Spectroscopy in MAS NMR of Biological and Organic Solids.
Spin Diffusion Driven by R-Symmetry Sequences: Applications to Homonuclear Correlation Spectroscopy in MAS NMR of Biological and Organic Solids.
Spin Diffusion Driven by R-Symmetry Sequences: Applications to Homonuclear Correlation Spectroscopy in MAS NMR of Biological and Organic Solids.
J Am Chem Soc. 2011 Mar 1;
Authors: Hou G, Yan S, Sun S, Han Y, Byeon IJ, Ahn J, Concel J, Samoson A, Gronenborn AM, Polenova T
We present a family of homonuclear (13)C-(13)Cmagic angle spinning spin diffusion experiments, based on R2(n)(v) (n = 1 and 2, v = 1...
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Spin Diffusion Driven by R-Symmetry Sequences: Applications to Homonuclear Correlation Spectroscopy in MAS NMR of Biological and Organic Solids
Spin Diffusion Driven by R-Symmetry Sequences: Applications to Homonuclear Correlation Spectroscopy in MAS NMR of Biological and Organic Solids
Guangjin Hou, Si Yan, Shangjin Sun, Yun Han, In-Ja L. Byeon, Jinwoo Ahn, Jason Concel, Ago Samoson, Angela M. Gronenborn and Tatyana Polenova
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Journal of the American Chemical Society
DOI: 10.1021/ja108650x
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[NMR paper] Slow diffusion of macromolecular assemblies by a new pulsed field gradient NMR method
Slow diffusion of macromolecular assemblies by a new pulsed field gradient NMR method.
Related Articles Slow diffusion of macromolecular assemblies by a new pulsed field gradient NMR method.
J Am Chem Soc. 2003 Mar 5;125(9):2541-5
Authors: Ferrage F, Zoonens M, Warschawski DE, Popot JL, Bodenhausen G
The translational diffusion coefficient of an integral membrane protein/surfactant complex has been measured using a novel pulsed field gradient NMR method. In this new approach, the information about the localization of the molecules is...
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[NMR paper] Practical methods for solid-state NMR distance measurements on large biomolecules: co
Practical methods for solid-state NMR distance measurements on large biomolecules: constant-time rotational resonance.
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J Magn Reson. 1999 Aug;139(2):371-6
Authors: Balazs YS, Thompson LK
Simple modifications of the rotational resonance experiment substantially reduce the total experimental time needed to measure weak homonuclear dipolar couplings, a critical factor for achieving routine internuclear distance...
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Using NMR to study fast dynamics in proteins: methods and applications.
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Curr Opin Pharmacol. 2010 Oct 6;
Authors: Sapienza PJ, Lee AL
Proteins exist not as singular structures with precise coordinates, but rather as fluctuating bodies that move rapidly through an enormous number of conformational substates. These dynamics have important implications for understanding protein function and for structure-based drug design. NMR spectroscopy is particularly well...
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[NMR paper] The hydration of proteins in solutions by self-diffusion coefficients. NMR study.
The hydration of proteins in solutions by self-diffusion coefficients. NMR study.
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Biochim Biophys Acta. 1996 Apr 17;1289(3):312-4
Authors: Baranowska HM, Olszewski KJ
The hydration of the globular (lysozyme, albumin) and fibrillar (fibrinogen) proteins in solution has been determined from the measurements of the self-diffusion coefficient by NMR pulsed gradient method. It has been concluded that the concentration dependencies of the...
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Using NMR Spectroscopic Methods to Determine Enantiomeric Purity and Assign Absolute
Using NMR Spectroscopic Methods to Determine Enantiomeric Purity and Assign Absolute Stereochemistry
Publication year: 2010
Source: Progress in Nuclear Magnetic Resonance Spectroscopy, In Press, Accepted Manuscript, Available online 2 August 2010</br>
Thomas J., Wenzel , Cora D., Chisholm</br>
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Methods to determine slow diffusion coefficients of biomolecules. Applications to Engrailed 2, a partially disordered protein
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