Abstract
A major goal in structural biology is to unravel how molecular machines function in detail. To that end, solution-state NMR spectroscopy is ideally suited as it is able to study biological assemblies in a near natural environment. Based on methyl TROSY methods, it is now possible to record high-quality data on complexes that are far over 100 kDa in molecular weight. In this review, we discuss the theoretical background of methyl TROSY spectroscopy, the information that can be extracted from methyl TROSY spectra and approaches that can be used to assign methyl resonances in large complexes. In addition, we touch upon insights that have been obtained for a number of challenging biological systems, including the 20S proteasome, the RNA exosome, molecular chaperones and G-protein-coupled receptors. We anticipate that methyl TROSY methods will be increasingly important in modern structural biology approaches, where information regarding static structures is complemented with insights into conformational changes and dynamic intermolecular interactions.
[NMR paper] Erratum to: Methyl labeling and TROSY NMR spectroscopy of proteins expressed in the eukaryote Pichia pastoris.
Erratum to: Methyl labeling and TROSY NMR spectroscopy of proteins expressed in the eukaryote Pichia pastoris.
Related Articles Erratum to: Methyl labeling and TROSY NMR spectroscopy of proteins expressed in the eukaryote Pichia pastoris.
J Biomol NMR. 2016 Feb 19;
Authors: Clark L, Zahm JA, Ali R, Kukula M, Bian L, Patrie SM, Gardner KH, Rosen MK, Rosenbaum DM
PMID: 26894386
Magic Angle Spinning NMR Spectroscopy: A Versatile Technique for Structural and Dynamic Analysis of Solid-Phase Systems
From The DNP-NMR Blog:
Magic Angle Spinning NMR Spectroscopy: A Versatile Technique for Structural and Dynamic Analysis of Solid-Phase Systems
This review gives a comprehensive overview of the state-of-the-art of magic-angle spinning (MAS), solid-state NMR spectroscopy, including DNP-NMR spectroscopy.
1. Polenova, T., R. Gupta, and A. Goldbourt, Magic angle spinning NMR spectroscopy: a versatile technique for structural and dynamic analysis of solid-phase systems. Anal Chem, 2015. 87(11): p. 5458-69.
nmrlearner
News from NMR blogs
0
06-29-2015 07:21 PM
[NMR paper] Methyl labeling and TROSY NMR spectroscopy of proteins expressed in the eukaryote Pichia pastoris.
Methyl labeling and TROSY NMR spectroscopy of proteins expressed in the eukaryote Pichia pastoris.
Related Articles Methyl labeling and TROSY NMR spectroscopy of proteins expressed in the eukaryote Pichia pastoris.
J Biomol NMR. 2015 May 30;
Authors: Clark L, Zahm JA, Ali R, Kukula M, Bian L, Patrie SM, Gardner KH, Rosen MK, Rosenbaum DM
Abstract
(13)C Methyl TROSY NMR spectroscopy has emerged as a powerful method for studying the dynamics of large systems such as macromolecular assemblies and membrane proteins. Specific...
nmrlearner
Journal club
0
05-31-2015 01:57 PM
Methyl labeling and TROSY NMR spectroscopy of proteins expressed in the eukaryote Pichia pastoris
Methyl labeling and TROSY NMR spectroscopy of proteins expressed in the eukaryote Pichia pastoris
Abstract
13C Methyl TROSY NMR spectroscopy has emerged as a powerful method for studying the dynamics of large systems such as macromolecular assemblies and membrane proteins. Specific 13C labeling of aliphatic methyl groups and perdeuteration has been limited primarily to proteins expressed in E. coli, preventing studies of many eukaryotic proteins of physiological and biomedical significance. We demonstrate the feasibility of...
nmrlearner
Journal club
0
05-29-2015 01:24 PM
Solution NMR spectroscopy of supra-molecular systems, why bother? A methyl-TROSY view.
Solution NMR spectroscopy of supra-molecular systems, why bother? A methyl-TROSY view.
Solution NMR spectroscopy of supra-molecular systems, why bother? A methyl-TROSY view.
J Magn Reson. 2011 Mar 30;
Authors: Kay LE
With the development of appropriate labeling schemes and the associated experiments that exploit them it has become possible to record high quality solution NMR spectra of supra-molecular complexes with molecular masses extending to 1MDa. One such approach involves selective (13)CH(3) methyl labeling in highly deuterated proteins...
nmrlearner
Journal club
0
04-05-2011 10:22 PM
DEER in Biological Multispin-Systems: A Case Study on the Fatty Acid Binding to Human Serum Albumin
DEER in Biological Multispin-Systems: A Case Study on the Fatty Acid Binding to Human Serum Albumin
Publication year: 2011
Source: Journal of Magnetic Resonance, In Press, Accepted Manuscript, Available online 10 March 2011</br>
Matthias J.N., Junk , Hans W., Spiess , Dariush, Hinderberger</br>
In this study, self-assembled systems of human serum albumin (HSA) and spin-labeled fatty acids are characterized by double electron–electron resonance (DEER). HSA, being the most important transport protein of the human blood, is capable to host up to seven paramagnetic fatty acid...
nmrlearner
Journal club
0
03-11-2011 05:00 PM
Solution NMR Spectroscopy of Supra-Molecular Systems, Why Bother? A Methyl TROSY View
Solution NMR Spectroscopy of Supra-Molecular Systems, Why Bother? A Methyl TROSY View
Publication year: 2011
Source: Journal of Magnetic Resonance, In Press, Accepted Manuscript, Available online 8 March 2011</br>
Lewis E., Kay</br>
With the development of appropriate labeling schemes and the associated experiments that exploit them it has become possible to record high quality solution NMR spectra of supra-molecular complexes with molecular masses extending to 1MDa. One such approach involves selective 13CH3 methyl labeling in highly deuterated proteins using experiments that make...
Methyl TROSY spectroscopy: A versatile NMR approach to study challenging biological systems.
Linear Mode
