[NMR paper] Enabling NMR studies of high molecular weight systems without the need for deuteration: The XL-ALSOFAST experiment with delayed decoupling.
Related ArticlesEnabling NMR studies of high molecular weight systems without the need for deuteration: The XL-ALSOFAST experiment with delayed decoupling.
Angew Chem Int Ed Engl. 2020 Aug 03;:
Authors: Rößler P, Mathieu D, Gossert AD
Abstract
Current biological research increasingly focusses on large human proteins and their complexes. Such proteins are difficult to study by NMR spectroscopy because they often can only be produced in higher eukaryotic expression systems, where deuteration is hardly feasible. Here, we present the XL-ALSOFAST-[ 13 C, 1 H]-HMQC experiment with much improved sensitivity for fully protonated high-molecular weight proteins. For the tested systems ranging from 100 to 240*kDa in size, 3-fold higher sensitivity was obtained on average for fast relaxing signals compared to current state-of-the-art experiments. In the XL-ALSOFAST approach, non-observed magnetisation is optimally exploited and transverse relaxation is minimized by the newly introduced concept of delayed decoupling. The combination of high sensitivity and superior artefact suppression makes it ideal for studying inherently unstable membrane proteins or for analysing therapeutic antibodies at natural 13 C abundance. The XL-ALSOFAST and delayed decoupling will therefore expand the range of biomolecular systems accessible to NMR spectroscopy.
PMID: 32743971 [PubMed - as supplied by publisher]
[NMR paper] A methyl-TROSY approach for NMR studies of high-molecular-weight DNA with application to the nucleosome core particle.
A methyl-TROSY approach for NMR studies of high-molecular-weight DNA with application to the nucleosome core particle.
Related Articles A methyl-TROSY approach for NMR studies of high-molecular-weight DNA with application to the nucleosome core particle.
Proc Natl Acad Sci U S A. 2020 May 26;:
Authors: Abramov G, Velyvis A, Rennella E, Wong LE, Kay LE
Abstract
The development of methyl-transverse relaxation-optimized spectroscopy (methyl-TROSY)-based NMR methods, in concert with robust strategies for incorporation of methyl-group...
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Application Note: Isotope Labeling of Alanine Methyl Groups for NMR Studies of High-Molecular-Weight Proteins - SelectScience
Application Note: Isotope Labeling of Alanine Methyl Groups for NMR Studies of High-Molecular-Weight Proteins - SelectScience
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Application Note: Isotope Labeling of Alanine Methyl Groups for NMR Studies of High-Molecular-Weight Proteins
SelectScience
Labeling schemes commonly employed for NMR investigations of high-molecular-weight proteins utilize selective incorporation of protons and 13C isotopes into methyl groups of Ileδ1, Leuδ and Valγ side-chains in a highly deuterated environment (commonly ...
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[NMR paper] (13)CHD 2-CEST NMR spectroscopy provides an avenue for studies of conformational exchange in high molecular weight proteins.
(13)CHD 2-CEST NMR spectroscopy provides an avenue for studies of conformational exchange in high molecular weight proteins.
Related Articles (13)CHD 2-CEST NMR spectroscopy provides an avenue for studies of conformational exchange in high molecular weight proteins.
J Biomol NMR. 2015 Aug 14;
Authors: Rennella E, Huang R, Velyvis A, Kay LE
Abstract
An NMR experiment for quantifying slow (millisecond) time-scale exchange processes involving the interconversion between visible ground state and invisible, conformationally excited...
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08-15-2015 04:01 PM
13 CHD 2 â??CEST NMR spectroscopy provides an avenue for studies of conformational exchange in high molecular weight proteins
13 CHD 2 â??CEST NMR spectroscopy provides an avenue for studies of conformational exchange in high molecular weight proteins
Abstract
An NMR experiment for quantifying slow (millisecond) time-scale exchange processes involving the interconversion between visible ground state and invisible, conformationally excited state conformers is presented. The approach exploits chemical exchange saturation transfer (CEST) and makes use of 13CHD2 methyl group probes that can be readily incorporated into otherwise highly deuterated proteins. The methodology is...
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08-14-2015 10:12 PM
O-tert-Butyltyrosine, an NMR Tag for High-Molecular-Weight Systems and Measurements of Submicromolar Ligand Binding Affinities
O-tert-Butyltyrosine, an NMR Tag for High-Molecular-Weight Systems and Measurements of Submicromolar Ligand Binding Affinities
Wan-Na Chen, Kekini Vahini Kuppan, Michael David Lee, Kristaps Jaudzems, Thomas Huber and Gottfried Otting
http://pubs.acs.org/appl/literatum/publisher/achs/journals/content/jacsat/0/jacsat.ahead-of-print/jacs.5b01918/20150327/images/medium/ja-2015-01918z_0002.gif
Journal of the American Chemical Society
DOI: 10.1021/jacs.5b01918
http://feeds.feedburner.com/~ff/acs/jacsat?d=yIl2AUoC8zA ...
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[NMR paper] O-tert-Butyltyrosine, an NMR tag for high-molecular weight systems and measurements of submicromolar ligand binding affinities.
O-tert-Butyltyrosine, an NMR tag for high-molecular weight systems and measurements of submicromolar ligand binding affinities.
O-tert-Butyltyrosine, an NMR tag for high-molecular weight systems and measurements of submicromolar ligand binding affinities.
J Am Chem Soc. 2015 Mar 19;
Authors: Chen WN, Kuppan KV, Lee MD, Jaudzems K, Huber T, Otting G
Abstract
O-tert-butyl-tyrosine (Tby) is an unnatural amino acid that can be site-specifically incorporated into proteins using established orthogonal aminoacyl-tRNA synthetase/tRNA...
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[NMR paper] Specific labeling and assignment strategies of valine methyl groups for NMR studies of high molecular weight proteins.
Specific labeling and assignment strategies of valine methyl groups for NMR studies of high molecular weight proteins.
http://www.bionmr.com//www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--production.springer.de-OnlineResources-Logos-springerlink.gif Related Articles Specific labeling and assignment strategies of valine methyl groups for NMR studies of high molecular weight proteins.
J Biomol NMR. 2013 Sep 28;
Authors: Mas G, Crublet E, Hamelin O, Gans P, Boisbouvier J
Abstract
The specific protonation of valine and leucine methyl...
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10-01-2013 11:15 PM
[NMR paper] Quantitative NMR studies of high molecular weight proteins: application to domain ori
Quantitative NMR studies of high molecular weight proteins: application to domain orientation and ligand binding in the 723 residue enzyme malate synthase G.
Related Articles Quantitative NMR studies of high molecular weight proteins: application to domain orientation and ligand binding in the 723 residue enzyme malate synthase G.
J Mol Biol. 2003 Apr 11;327(5):1121-33
Authors: Tugarinov V, Kay LE
A high-resolution multidimensional NMR study of ligand-binding to Escherichia coli malate synthase G (MSG), a 723-residue monomeric enzyme (81.4...
Enabling NMR studies of high molecular weight systems without the need for deuteration: The XL-ALSOFAST experiment with delayed decoupling.
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