A solubilizing "Trt-K10" tag was developed for the effective chemical preparation of peptides/proteins with low solubility. The Trt-K10 tag comprises a hydrophilic oligo-Lys sequence and a trityl anchor, and can be selectively introduced to a side chain thiol of Cys of deprotected peptides/proteins with a trityl alcohol-type introducing reagent "Trt(OH)-K10" under acidic conditions. Significantly, the ligation product in the reaction mixture of a thiol-additive-free native chemical ligation can be modified directly in a one-pot manner to facilitate the isolation of the product by high-performance liquid chromatography. Finally, the Trt-K10 tag can be readily removed with a standard trifluoroacetic acid cocktail. Using this easy-to-attach/detach tag-aided method, a hepatitis B virus capsid protein that is usually difficult to handle was synthesized successfully.
[NMR paper] P-B desulfurization: an enabling method for protein chemical synthesis and site-specific deuteration
P-B desulfurization: an enabling method for protein chemical synthesis and site-specific deuteration
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...
nmrlearner
Journal club
0
10-03-2017 03:24 AM
[NMR paper] Chemical Synthesis of the Highly Hydrophobic Antiviral Membrane Associated Protein IFITM3 and Modified Variants
Chemical Synthesis of the Highly Hydrophobic Antiviral Membrane Associated Protein IFITM3 and Modified Variants
Interferon-induced transmembrane protein 3 (IFITM3) is an antiviral transmembrane protein thought to serve as the primary factor for inhibiting replication of a large number of viruses, including the West Nile virus, the Dengue virus, the Ebola virus, and the Zika virus. Production of this 14.5 kDa, 133-residue, transmembrane protein, especially with essential posttranslational modifications by recombinant expression is challenging. In this report, we document the chemical...
nmrlearner
Journal club
0
08-21-2017 02:23 PM
[NMR paper] Palladium in Chemical Protein Synthesis and Modifications
Palladium in Chemical Protein Synthesis and Modifications
The field of site-specific modification of proteins has drawn significant attentions in recent years owing to its high importance in various research areas such as the development of novel therapeutics and understanding the biochemical and cellular behaviors of proteins. The presence of a large number of reactive functional groups in the protein of interest and in the cellular environment renders the particular modification at a specific site a highly challenging task. However, with the development of sophisticated...
[NMR paper] The Use of Amphipols for Solution NMR Studies of Membrane Proteins: Advantages and Constraints as Compared to Other Solubilizing Media.
The Use of Amphipols for Solution NMR Studies of Membrane Proteins: Advantages and Constraints as Compared to Other Solubilizing Media.
Related Articles The Use of Amphipols for Solution NMR Studies of Membrane Proteins: Advantages and Constraints as Compared to Other Solubilizing Media.
J Membr Biol. 2014 Mar 28;
Authors: Planchard N, Point E, Dahmane T, Giusti F, Renault M, Le Bon C, Durand G, Milon A, Guittet E, Zoonens M, Popot JL, Catoire LJ
Abstract
Solution-state nuclear magnetic resonance studies of membrane proteins are...
nmrlearner
Journal club
0
03-29-2014 01:00 PM
[NMR paper] CAMRA: chemical shift based computer aided protein NMR assignments.
CAMRA: chemical shift based computer aided protein NMR assignments.
Related Articles CAMRA: chemical shift based computer aided protein NMR assignments.
J Biomol NMR. 1998 Oct;12(3):395-405
Authors: Gronwald W, Willard L, Jellard T, Boyko RF, Rajarathnam K, Wishart DS, Sönnichsen FD, Sykes BD
A suite of programs called CAMRA (Computer Aided Magnetic Resonance Assignment) has been developed for computer assisted residue-specific assignments of proteins. CAMRA consists of three units: ORB, CAPTURE and PROCESS. ORB predicts NMR chemical shifts...
nmrlearner
Journal club
0
11-17-2010 11:15 PM
[NMR paper] Posttranslational modification of Klebsiella pneumoniae flavodoxin by covalent attach
Posttranslational modification of Klebsiella pneumoniae flavodoxin by covalent attachment of coenzyme A, shown by 31P NMR and electrospray mass spectrometry, prevents electron transfer from the nifJ protein to nitrogenase. A possible new regulatory mechanism for biological nitrogen fixation.
Related Articles Posttranslational modification of Klebsiella pneumoniae flavodoxin by covalent attachment of coenzyme A, shown by 31P NMR and electrospray mass spectrometry, prevents electron transfer from the nifJ protein to nitrogenase. A possible new regulatory mechanism for biological...
nmrlearner
Journal club
0
08-21-2010 11:41 PM
[NMR paper] Computer-aided assignment of the 1H-NMR spectrum of the viral-protein-genome-linked p
Computer-aided assignment of the 1H-NMR spectrum of the viral-protein-genome-linked polypeptide from cowpea mosaic virus.
http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--www3.interscience.wiley.com-aboutus-images-wiley_interscience_pubmed_logo_FREE_120x27.gif Related Articles Computer-aided assignment of the 1H-NMR spectrum of the viral-protein-genome-linked polypeptide from cowpea mosaic virus.
Eur J Biochem. 1990 Jul 5;190(3):583-91
Authors: van de Ven FJ, Lycksell PO, van Kammen A, Hilbers CW
The 1H-NMR spectrum of the...
Easy-to-Attach/Detach Solubilizing Tag-Aided Chemical Synthesis of an Aggregative Capsid Protein
Linear Mode
