Native chemical ligation has provided a powerful tool for protein chemical synthesis. Herein, we report an unprecedented mild system (TCEP-NaBH4 or TCEP-LiBEt3H) for chemoselective peptide desulfurization to achieve effective protein synthesis via the native chemical ligation-desulfurization approach. This method, termed P-B desulfurization, features usage of common reagents, simplicity of operation, robustness, high yielding, clean conversion and versatile functionality compatibility with complex peptides/proteins. In addition, this method allows for incorporating deuterium into the peptides after cysteine desulfurization when running the reaction in D2O buffer. Moreover, this method enables clean desulfurization for peptides carrying post-translational modifications, such as phosphorylation and crotonylation. The effectiveness of this method has been demonstrated in the synthesis of cyclic peptides Dichotomin, and synthetic proteins including ubiquitin, ?-synuclein and histone H2A.
[NMR paper] Erratum to: Protein-protein interaction analysis in crude bacterial lysates using combinational method of (19)F site-specific incorporation and (19)F NMR.
Erratum to: Protein-protein interaction analysis in crude bacterial lysates using combinational method of (19)F site-specific incorporation and (19)F NMR.
Related Articles Erratum to: Protein-protein interaction analysis in crude bacterial lysates using combinational method of (19)F site-specific incorporation and (19)F NMR.
Protein Cell. 2017 Mar 10;:
Authors: Li D, Zhang Y, He Y, Zhang C, Wang J, Xiong Y, Zhang L, Liu Y, Shi P, Tian C
PMID: 28284007
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[NMR paper] Chemo-enzymatic synthesis of site-specific isotopically labeled nucleotides for use in NMR resonance assignment, dynamics and structural characterizations.
Chemo-enzymatic synthesis of site-specific isotopically labeled nucleotides for use in NMR resonance assignment, dynamics and structural characterizations.
http://www.bionmr.com//www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--highwire.stanford.edu-icons-externalservices-pubmed-oup_oa.gif http://www.bionmr.com//www.ncbi.nlm.nih.gov/corehtml/query/egifs/https:--www.ncbi.nlm.nih.gov-corehtml-pmc-pmcgifs-pubmed-pmc.gif Related Articles Chemo-enzymatic synthesis of site-specific isotopically labeled nucleotides for use in NMR resonance assignment, dynamics and structural...
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08-31-2016 02:34 PM
[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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A chemical approach for site-specific identification of NMR signals from protein side-chain NH 3 + groups forming intermolecular ion pairs in proteinā??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 proteinā??nucleic acid complexes
Abstract
Proteinā??nucleic acid interactions involve intermolecular ion pairs of protein side-chain and DNA or RNA phosphate groups. Using three proteinā??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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[NMR paper] A Magic-Angle Spinning NMR Method for the Site-Specific Measurement of Proton Chemical-Shift Anisotropy in Biological and Organic Solids.
A Magic-Angle Spinning NMR Method for the Site-Specific Measurement of Proton Chemical-Shift Anisotropy in Biological and Organic Solids.
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Isr J Chem. 2014 Feb 1;54(1-2):171-183
Authors: Hou G, Gupta R, Polenova T, Vega AJ
Abstract
Proton chemical shifts are a rich probe of structure and hydrogen bonding environments in organic and biological molecules. Until recently,...
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[NMR paper] Preparation of Multiple Site-Specific Mutant Proteins for NMR Studies by PCR-Directed Cell-Free Protein Synthesis.
Preparation of Multiple Site-Specific Mutant Proteins for NMR Studies by PCR-Directed Cell-Free Protein Synthesis.
Preparation of Multiple Site-Specific Mutant Proteins for NMR Studies by PCR-Directed Cell-Free Protein Synthesis.
Methods Mol Biol. 2014;1118:169-87
Authors: Ozawa K, Qi R
Abstract
Cell-free protein synthesis (CFPS) offers a fast and inexpensive approach to selectively label proteins with isotopes that can then be detected by nuclear magnetic resonance (NMR) spectroscopy directly in the translation mixture. We describe...
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[NMR paper] Site-specific 13C chemical shift anisotropy measurements in a uniformly 15N,13C-labeled microcrystalline protein by 3D magic-angle spinning NMR spectroscopy.
Site-specific 13C chemical shift anisotropy measurements in a uniformly 15N,13C-labeled microcrystalline protein by 3D magic-angle spinning NMR spectroscopy.
Related Articles Site-specific 13C chemical shift anisotropy measurements in a uniformly 15N,13C-labeled microcrystalline protein by 3D magic-angle spinning NMR spectroscopy.
J Am Chem Soc. 2005 Aug 31;127(34):11946-7
Authors: Wylie BJ, Franks WT, Graesser DT, Rienstra CM
In this Communication, we introduce a 3D magic-angle spinning recoupling experiment that correlates chemical shift...
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Site-Specific Protein Backbone and Side-Chain NMR Chemical Shift and Relaxation Analy
Site-Specific Protein Backbone and Side-Chain NMR Chemical Shift and Relaxation Analysis of Human Vinexin SH3 Domain using a Genetically Encoded (15)N/(19)F-Labeled Unnatural Amino Acid.
Related Articles Site-Specific Protein Backbone and Side-Chain NMR Chemical Shift and Relaxation Analysis of Human Vinexin SH3 Domain using a Genetically Encoded (15)N/(19)F-Labeled Unnatural Amino Acid.
Biochem Biophys Res Commun. 2010 Oct 11;
Authors: Shi P, Xi Z, Wang H, Shi C, Xiong Y, Tian C
SH3 is a ubiquitous domain mediating protein-protein interactions....
P-B desulfurization: an enabling method for protein chemical synthesis and site-specific deuteration
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