17O NMR studies of yeast ubiquitin in aqueous solution and in the solid state.
Chembiochem. 2020 Oct 15;:
Authors: Lin B, Hung I, Gan Z, Chien PH, Spencer H, Smith S, Wu G
Abstract
We report a general method for amino acid-type specific 17 O-labeling of recombinant proteins in Escherichia coli. In particular, we have prepared several [1- 13 C, 17 O]-labeled yeast ubiquitin (Ub) samples including Ub-[1- 13 C, 17 O]Gly, Ub-[1- 13 C, 17 O]Tyr, and Ub-[1- 13 C, 17 O]Phe using the auxotrophic E. coli strain DL39 GlyA l DE3 ( asp C - tyr B - ilv E - gly A - l DE3). We have also produced Ub-[ h - 17 O]Tyr, where the phenolic group of Tyr59 is 17 O-labeled. We showed for the first time that 17 O NMR signals from protein terminal residues and sidechains can be readily detected in aqueous solution. We also reported solid-state 17 O NMR spectra for Ub-[1- 13 C, 17 O]Tyr and Ub-[1- 13 C, 17 O]Phe obtained at an ultrahigh magnetic field, 35.2 T (1.5 GHz for 1 H). This work represents a significant advance in the field of 17 O NMR studies of proteins.
PMID: 33058374 [PubMed - as supplied by publisher]
Uniform isotope labeling of a eukaryotic seven-transmembrane helical protein in yeast enables high-resolution solid-state NMR studies in the lipid environment
Uniform isotope labeling of a eukaryotic seven-transmembrane helical protein in yeast enables high-resolution solid-state NMR studies in the lipid environment
Abstract Overexpression of isotope-labeled multi-spanning eukaryotic membrane proteins for structural NMR studies is often challenging. On the one hand, difficulties with achieving proper folding, membrane insertion, and native-like post-translational modifications frequently disqualify bacterial expression systems. On the other hand, eukaryotic cell cultures can be prohibitively expensive. One of the viable alternatives,...
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Uniform isotope labeling of a eukaryotic seven-transmembrane helical protein in yeast enables high-resolution solid-state NMR studies in the lipid environment.
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17O NMR studies of yeast ubiquitin in aqueous solution and in the solid state.
Linear Mode
