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Site-selective C labeling of proteins using erythrose

J. Biomol. NMR. 2017-03; 
WeiningerUl
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PCR and Cloning … Expression and purification. An optimised coding sequence for human FK506 binding protein 12 (FKBP12; Uniprot: P62942) was synthesised (GenScript, Piscataway, NJ, USA) and sub-cloned into the plasmid pNIC28-Bsa4 (Savitsky et al. 2010) … Get A Quote

摘要

NMR-spectroscopy enables unique experimental studies on protein dynamics at atomic resolution. In order to obtain a full atom view on protein dynamics, and to study specific local processes like ring-flips, proton-transfer, or tautomerization, one has to perform studies on amino-acid side chains. A key requirement for these studies is site-selective labeling with C and/or H, which is achieved in the most general way by using site-selectively C-enriched glucose (1- and 2-C) as the carbon source in bacterial expression systems. Using this strategy, multiple sites in side chains, including aromatics, become site-selectively labeled and suitable for relaxation studies. Here we systematically investi... More

关键词

Aromatic side chain,Isotope labeling,Protein dynamics,Relaxa