Publication date: Available online 26 June 2018 Source:Methods
Author(s): Scott D. Gorman, Debashish Sahu, Kathleen F. O'Rourke, David D. Boehr
Solution-state NMR is an important tool for studying protein structure and function. The ability to probe methyl groups has helped to substantially expand the scope of proteins accessible by NMR spectroscopy, including facilitating study of proteins and complexes greater than 100 kDa in size. While the toolset for studying protein structure and dynamics by NMR continues to expand, a major rate-limiting step in these studies is the initial resonance assignments, especially for larger (> 50 kDa) proteins. In this practical review, we present strategies for efficiently isotopically labeling proteins, delineate NMR pulse sequences that can be used to determine methyl resonance assignments in the presence and absence of backbone assignments, and outline computational methods for NMR data analysis. We use our experiences from assigning methyl resonances for the aromatic biosynthetic enzymes tryptophan synthase and chorismate mutase to provide advice for all stages of experimental set-up and data analysis.
[NMR paper] Assignment of methyl NMR resonances of a 52*kDa protein with residue-specific 4D correlation maps.
Assignment of methyl NMR resonances of a 52*kDa protein with residue-specific 4D correlation maps.
Related Articles Assignment of methyl NMR resonances of a 52*kDa protein with residue-specific 4D correlation maps.
J Biomol NMR. 2015 May 8;
Authors: Mishra SH, Frueh DP
Abstract
Methyl groups have become key probes for structural and functional studies by nuclear magnetic resonance. However, their NMR signals cluster in a small spectral region and assigning their resonances can be a tedious process. Here, we present a method...
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Assignment of methyl NMR resonances of a 52Â*kDa protein with residue-specific 4D correlation maps
Assignment of methyl NMR resonances of a 52Â*kDa protein with residue-specific 4D correlation maps
Abstract
Methyl groups have become key probes for structural and functional studies by nuclear magnetic resonance. However, their NMR signals cluster in a small spectral region and assigning their resonances can be a tedious process. Here, we present a method that facilitates assignment of methyl resonances from assigned amide groups. Calculating the covariance between sensitive methyl and amide 3D spectra, each providing correlations to Cα and Cβ...
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05-07-2015 03:04 PM
Methyl-specific isotopic labeling: a molecular tool box for solution NMR studies of large proteins
Methyl-specific isotopic labeling: a molecular tool box for solution NMR studies of large proteins
Publication date: June 2015
Source:Current Opinion in Structural Biology, Volume 32</br>
Author(s): Rime Kerfah , Michael J Plevin , Remy Sounier , Pierre Gans , Jerome Boisbouvier</br>
Nuclear magnetic resonance (NMR) spectroscopy is a uniquely powerful tool for studying the structure, dynamics and interactions of biomolecules at atomic resolution. In the past 15 years, the development of new isotopic labeling strategies has opened the possibility of exploiting...
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04-14-2015 01:24 PM
[NMR paper] Specific (13)C labeling of leucine, valine and isoleucine methyl groups for unambiguous detection of long-range restraints in protein solid-state NMR studies.
Specific (13)C labeling of leucine, valine and isoleucine methyl groups for unambiguous detection of long-range restraints in protein solid-state NMR studies.
Specific (13)C labeling of leucine, valine and isoleucine methyl groups for unambiguous detection of long-range restraints in protein solid-state NMR studies.
J Magn Reson. 2015 Jan 6;252C:10-19
Authors: Fasshuber HK, Demers JP, Chevelkov V, Giller K, Becker S, Lange A
Abstract
Here we present an isotopic labeling strategy to easily obtain unambiguous long-range...
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01-28-2015 05:28 PM
Specific 13C labeling of leucine, valine and isoleucine methyl groups for unambiguous detection of long-range restraints in protein solid-state NMR studies
Specific 13C labeling of leucine, valine and isoleucine methyl groups for unambiguous detection of long-range restraints in protein solid-state NMR studies
Publication date: Available online 6 January 2015
Source:Journal of Magnetic Resonance</br>
Author(s): Hannes Klaus Fasshuber , Jean-Philippe Demers , Veniamin Chevelkov , Karin Giller , Stefan Becker , Adam Lange</br>
Here we present an isotopic labeling strategy to easily obtain unambiguous long-range distance restraints in protein solid-state NMR studies. The method is based on the inclusion of two...
[NMR paper] Stereospecific assignments of the leucine methyl resonances in the 1H NMR spectrum of
Stereospecific assignments of the leucine methyl resonances in the 1H NMR spectrum of Lactobacillus casei dihydrofolate reductase.
http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--linkinghub.elsevier.com-ihub-images-PubMedLink.gif Related Articles Stereospecific assignments of the leucine methyl resonances in the 1H NMR spectrum of Lactobacillus casei dihydrofolate reductase.
FEBS Lett. 1993 Mar 1;318(2):177-80
Authors: Ostler G, Soteriou A, Moody CM, Khan JA, Birdsall B, Carr MD, Young DW, Feeney J
A general method is described for the...
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[NMR paper] Solution studies of staphylococcal nuclease H124L. 1. Backbone 1H and 15N resonances
Solution studies of staphylococcal nuclease H124L. 1. Backbone 1H and 15N resonances and secondary structure of the unligated enzyme as identified by three-dimensional NMR spectroscopy.
Related Articles Solution studies of staphylococcal nuclease H124L. 1. Backbone 1H and 15N resonances and secondary structure of the unligated enzyme as identified by three-dimensional NMR spectroscopy.
Biochemistry. 1992 Jan 28;31(3):911-20
Authors: Wang JF, Mooberry ES, Walkenhorst WF, Markley JL
The backbone 1H and 15N resonances of unligated staphylococcal...
Assigning methyl resonances for protein solution-state NMR studies
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