Related ArticlesCell-Free Protein Synthesis of Small Intrinsically Disordered Proteins for NMR Spectroscopy.
Methods Mol Biol. 2020;2141:233-245
Authors: Isaksson L, Pedersen A
Abstract
Cell-free protein synthesis (CFPS) is an established method to produce recombinant proteins and has been used in a wide variety of applications. The use of CFPS has almost from the onset been favorably linked to the production of isotopically labelled proteins for NMR spectroscopy as the resulting labelling of the produced protein is defined by the chosen amino acids during reaction setup. Here we describe how to set up production and isotopic labelling of small intrinsically disordered proteins (IDPs) for NMR spectroscopy applications using an E. coli-based CFPS system in batch mode.
[NMR paper] Protein labeling strategies for liquid-state NMR spectroscopy using cell-free synthesis.
Protein labeling strategies for liquid-state NMR spectroscopy using cell-free synthesis.
Related Articles Protein labeling strategies for liquid-state NMR spectroscopy using cell-free synthesis.
Prog Nucl Magn Reson Spectrosc. 2018 Apr;105:1-22
Authors: Hoffmann B, Löhr F, Laguerre A, Bernhard F, Dötsch V
Abstract
Preparation of a protein sample for liquid-state nuclear magnetic resonance (NMR) spectroscopy analysis requires optimization of many parameters. This review describes labeling strategies for obtaining assignments of...
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03-20-2018 10:11 PM
Protein labeling strategies for liquid-state NMR spectroscopy using cell-free synthesis
Protein labeling strategies for liquid-state NMR spectroscopy using cell-free synthesis
Publication date: Available online 7 December 2017
Source:Progress in Nuclear Magnetic Resonance Spectroscopy</br>
Author(s): Beate Hoffmann, Frank Löhr, Aisha Laguerre, Frank Bernhard, Volker Dötsch</br>
Preparation of a protein sample for liquid-state nuclear magnetic resonance (NMR) spectroscopy analysis requires optimization of many parameters. This review describes labeling strategies for obtaining assignments of protein resonances. Particular emphasis is placed on...
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12-09-2017 07:49 AM
[NMR paper] In-cell (13)C NMR spectroscopy for the study of intrinsically disordered proteins.
In-cell (13)C NMR spectroscopy for the study of intrinsically disordered proteins.
In-cell (13)C NMR spectroscopy for the study of intrinsically disordered proteins.
Nat Protoc. 2014 Sep;9(9):2005-2016
Authors: Felli IC, Gonnelli L, Pierattelli R
Abstract
A large number of proteins carry out their function in highly flexible and disordered states, lacking a well-defined 3D structure. These proteins, referred to as intrinsically disordered proteins (IDPs), are now in the spotlight of modern structural biology. Nuclear magnetic...
[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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01-08-2014 11:23 AM
[NMR paper] Cell-free synthesis of 15N-labeled proteins for NMR studies.
Cell-free synthesis of 15N-labeled proteins for NMR studies.
Related Articles Cell-free synthesis of 15N-labeled proteins for NMR studies.
IUBMB Life. 2005 Sep;57(9):615-22
Authors: Ozawa K, Dixon NE, Otting G
Modern cell-free in vitro protein synthesis systems present powerful tools for the synthesis of isotope-labeled proteins in high yields. The production of selectively 15 N-labeled proteins from 15 N-labeled amino acids is particularly economic and yields are often sufficient to analyze the proteins very quickly by two-dimensional NMR...
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12-01-2010 06:56 PM
Cell-free protein synthesis of perdeuterated proteins for NMR studies
Cell-free protein synthesis of perdeuterated proteins for NMR studies
Touraj Etezady-Esfarjani, Sebastian Hiller, Cristina Villalba and Kurt Wüthrich
Journal of Biomolecular NMR; 2007; 39(3); pp 229-238
Abstract:
Cell-free protein synthesis protocols for uniformly deuterated proteins typically yield low, non-uniform deuteration levels. This paper introduces an E. coli cell-extract, D-S30, which enables efficient production of proteins with high deuteration levels for all non-labile hydrogen atom positions. Potential applications of the new protocol may include production of proteins...
Cell-Free Protein Synthesis of Small Intrinsically Disordered Proteins for NMR Spectroscopy.
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