Using amber suppression in coordination with a mutant pyrrolysyl-tRNA synthetase-tRNAPyl pair, azidonorleucine is genetically encoded in E. coli. Its genetic incorporation followed by traceless Staudinger ligation with a phosphinothioester allows the convenient synthesis of a protein with a site-specifically installed lysine acylation. By simply changing the phosphinothioester identity, any lysine acylation type could be introduced. Using this approach, we demonstrated that both lysine acetylation and lysine succinylation can be installed selectively in ubiquitin and synthesized histone H3 with succinylation at its K4 position (H3K4su). Using an H3K4su-H4 tetramer as a substrate, we further confirmed that Sirt5 is an active histone desuccinylase. Lysine succinylation is a recently identified post-translational modification. The reported technique makes it possible to explicate regulatory functions of this modification in proteins.Azidonorleucine, an azide-containing amino acid, is genetically encoded and incorporated into model proteins. This incorporation followed by traceless Staudinger ligation potentiates the synthesis of proteins with a myriad of site-specific lysine acylations.
[NMR paper] A chemical approach for site-specific identification of NMR signals from protein side-chain NH3 (+) groups forming intermolecular ion pairs in protein-nucleic acid complexes.
A chemical approach for site-specific identification of NMR signals from protein side-chain NH3 (+) groups forming intermolecular ion pairs in protein-nucleic acid complexes.
Related Articles A chemical approach for site-specific identification of NMR signals from protein side-chain NH3 (+) groups forming intermolecular ion pairs in protein-nucleic acid complexes.
J Biomol NMR. 2015 Feb 19;
Authors: Anderson KM, Nguyen D, Esadze A, Zandrashvili L, Gorenstein DG, Iwahara J
Abstract
Protein-nucleic acid interactions involve...
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02-19-2015 07:03 PM
A chemical approach for site-specific identification of NMR signals from protein side-chain NH 3 + groups forming intermolecular ion pairs in proteinâ??nucleic acid complexes
A chemical approach for site-specific identification of NMR signals from protein side-chain NH 3 + groups forming intermolecular ion pairs in proteinâ??nucleic acid complexes
Abstract
Proteinâ??nucleic acid interactions involve intermolecular ion pairs of protein side-chain and DNA or RNA phosphate groups. Using three proteinâ??DNA complexes, we demonstrate that site-specific oxygen-to-sulfur substitution in phosphate groups allows for identification of NMR signals from the protein side-chain NH3 + groups forming the...
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02-18-2015 06:15 PM
Site-specific analysis of heteronuclear Overhauser effects in microcrystalline proteins
Site-specific analysis of heteronuclear Overhauser effects in microcrystalline proteins
Abstract
Relaxation parameters such as longitudinal relaxation are susceptible to artifacts such as spin diffusion, and can be affected by paramagnetic impurities as e.g. oxygen, which make a quantitative interpretation difficult. We present here the site-specific measurement of 13C and 15N heteronuclear rates in an immobilized protein. For methyls, a strong effect is expected due to the three-fold rotation of the methyl group. Quantification of the 13C...
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07-03-2014 06:04 AM
Site-specific dynamic nuclear polarization of hydration water as a generally applicable approach to monitor protein aggregation
From The DNP-NMR Blog:
Site-specific dynamic nuclear polarization of hydration water as a generally applicable approach to monitor protein aggregation
This article was already published in 2009 but unfortunately I missed it.
Pavlova, A., et al., Site-specific dynamic nuclear polarization of hydration water as a generally applicable approach to monitor protein aggregation. Phys. Chem. Chem. Phys., 2009. 11(31): p. 6833-6839.
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11-21-2013 01:14 AM
[NMR paper] Investigation of Lysine Side Chain Interactions of Interleukin-8 with Heparin and other Glycosaminoglycans Studied by a Methylation-NMR Approach.
Investigation of Lysine Side Chain Interactions of Interleukin-8 with Heparin and other Glycosaminoglycans Studied by a Methylation-NMR Approach.
Investigation of Lysine Side Chain Interactions of Interleukin-8 with Heparin and other Glycosaminoglycans Studied by a Methylation-NMR Approach.
Glycobiology. 2013 Aug 27;
Authors: Möbius K, Nordsieck K, Pichert A, Samsonov SA, Thomas L, Schiller J, Kalkhof S, Pisabarro MT, Beck-Sickinger AG, Huster D
Abstract
Although the interaction between interleukin-8 (IL-8) and glycosaminoglycans (GAG) is...
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08-29-2013 01:53 PM
Site-Specific Mapping and Time-Resolved Monitoring of Lysine Methylation by High-Resolution NMR Spectroscopy
Site-Specific Mapping and Time-Resolved Monitoring of Lysine Methylation by High-Resolution NMR Spectroscopy
Franc?ois-Xavier Theillet, Stamatios Liokatis, Jan Oliver Jost, Beata Bekei, Honor May Rose, Andres Binolfi, Dirk Schwarzer and Philipp Selenko
http://pubs.acs.org/appl/literatum/publisher/achs/journals/content/jacsat/0/jacsat.ahead-of-print/ja301895f/aop/images/medium/ja-2012-01895f_0003.gif
Journal of the American Chemical Society
DOI: 10.1021/ja301895f
http://feeds.feedburner.com/~ff/acs/jacsat?d=yIl2AUoC8zA
http://feeds.feedburner.com/~r/acs/jacsat/~4/tV2dPnMa4Qc
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04-27-2012 10:27 PM
Site-specific labeling of proteins with NMR-active unnatural amino acids
Site-specific labeling of proteins with NMR-active unnatural amino acids
Abstract A large number of amino acids other than the canonical amino acids can now be easily incorporated in vivo into proteins at genetically encoded positions. The technology requires an orthogonal tRNA/aminoacyl-tRNA synthetase pair specific for the unnatural amino acid that is added to the media while a TAG amber or frame shift codon specifies the incorporation site in the protein to be studied. These unnatural amino acids can be isotopically labeled and provide unique opportunities for site-specific labeling...
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01-09-2011 12:46 PM
[NMR paper] NMR analysis of site-specific ligand binding in oligomeric proteins. Dynamic studies
NMR analysis of site-specific ligand binding in oligomeric proteins. Dynamic studies on the interaction of riboflavin synthase with trifluoromethyl-substituted intermediates.
http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--pubs.acs.org-images-acspubs.jpg Related Articles NMR analysis of site-specific ligand binding in oligomeric proteins. Dynamic studies on the interaction of riboflavin synthase with trifluoromethyl-substituted intermediates.
Biochemistry. 1996 Jul 30;35(30):9637-46
Authors: Scheuring J, Fischer M, Cushman M, Lee J, Bacher A,...
A Versatile Approach for Site-Specific Lysine Acylation in Proteins
Linear Mode
