Cell-free protein synthesis of perdeuterated proteins for NMR studies

Etezady-Esfarjani, Touraj and Hiller, Sebastian and Villalba, Cristina and Wüthrich, Kurt. (2007) Cell-free protein synthesis of perdeuterated proteins for NMR studies. Journal of biomolecular NMR, 39 (3). pp. 229-238.

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Official URL: http://edoc.unibas.ch/41065/

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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 with selective isotope-labeling of selected amino acid residues on a perdeuterated background for studies of enzyme active sites or for ligand screening in drug discovery projects, as well as the synthesis of perdeuterated polypeptides for NMR spectroscopy with large supra-molecular structures. As an illustration, it is demonstrated that the 800-kDa chaperonine GroEL synthesized with the D-S30 cell-free system had a uniform deuteration level of about 95% and assembled into its biologically active oligomeric form.
Faculties and Departments:05 Faculty of Science > Departement Biozentrum > Structural Biology & Biophysics > Structural Biology (Hiller)
UniBasel Contributors:Hiller Odermatt, Sebastian
Item Type:Article, refereed
Article Subtype:Research Article
Note:Publication type according to Uni Basel Research Database: Journal article
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Last Modified:19 Dec 2017 12:24
Deposited On:17 Aug 2016 07:59

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