Cell-free protein synthesis of perdeuterated proteins for NMR studies Export

@article{citeulike:4637747, abstract = {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 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.}, author = {Etezady-Esfarjani, Touraj and Hiller, Sebastian and Villalba, Cristina and W\"{u}thrich, Kurt}, citeulike-article-id = {4637747}, citeulike-linkout-0 = {http://dx.doi.org/10.1007/s10858-007-9188-0}, citeulike-linkout-1 = {http://www.springerlink.com/content/e36451521822lk84}, day = {1}, doi = {10.1007/s10858-007-9188-0}, journal = {Journal of Biomolecular NMR}, keywords = {biggies, cell-free, labeling}, month = {November}, number = {3}, pages = {229--238}, posted-at = {2009-05-26 22:53:04}, priority = {2}, title = {Cell-free protein synthesis of perdeuterated proteins for NMR studies}, url = {http://dx.doi.org/10.1007/s10858-007-9188-0}, volume = {39}, year = {2007} }

TY - JOUR ID - citeulike:4637747 L3 - citeulike-article-id:4637747 N2 - 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 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. IS - 3 JF - Journal of Biomolecular NMR EP - 238 TI - Cell-free protein synthesis of perdeuterated proteins for NMR studies VL - 39 SP - 229 KW - biggies KW - cell-free KW - labeling AU - Etezady-Esfarjani, Touraj AU - Hiller, Sebastian AU - Villalba, Cristina AU - Wüthrich, Kurt PY - 2007/11/01/ UR - http://dx.doi.org/10.1007/s10858-007-9188-0 ER -