ArticleJournal of biomolecular NMR2003
A novel method for the biosynthesis of deuterated proteins with selective protonation at the aromatic rings of Phe, Tyr and Trp.
Article in Journal of biomolecular NMR, 2003. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 13 papers.
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Who cites it
13 citing papers in PubMed.
- Site-Specific Isotope Labeling of FliG for Studying Structural Dynamics Using Nuclear Magnetic Resonance Spectroscopy.Methods in molecular biology (Clifton, N.J.) · 2023Article
- Specific isotopic labelling and reverse labelling for protein NMR spectroscopy: using metabolic precursors in sample preparation.Biochemical Society transactions · 2022Review
- Late metabolic precursors for selective aromatic residue labeling.Journal of biomolecular NMR · 2018Review
- Anthranilic acid, the new player in the ensemble of aromatic residue labeling precursor compounds.Journal of biomolecular NMR · 2017Article
- Bacterial production of site specificJournal of biomolecular NMR · 2017Article
- Highly efficient residue-selective labeling with isotope-labeled Ile, Leu, and Val using a new auxotrophic E. coli strain.Journal of biomolecular NMR · 2016Article
- The pentameric nucleoplasmin fold is present in Drosophila FKBP39 and a large number of chromatin-related proteins.Journal of molecular biology · 2015Article
- α-Ketoacids as precursors for phenylalanine and tyrosine labelling in cell-based protein overexpression.Journal of biomolecular NMR · 2013Article
- Efficient acquisition of high-resolution 4-D diagonal-suppressed methyl-methyl NOESY for large proteins.Journal of magnetic resonance (San Diego, Calif. : 1997) · 2012Article
- Cell-free synthesis of membrane subunits of ATP synthase in phospholipid bicelles: NMR shows subunit a fold similar to the protein in the cell membrane.Protein science : a publication of the Protein Society · 2012Article
- Alternative SAIL-Trp for robust aromatic signal assignment and determination of the χ(2) conformation by intra-residue NOEs.Journal of biomolecular NMR · 2011Article
- Efficient condensed-phase production of perdeuterated soluble and membrane proteins.Journal of structural and functional genomics · 2010Article
- Expression of human peripheral cannabinoid receptor for structural studies.Protein science : a publication of the Protein Society · 2005Article
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Authors and funding
7 authors.
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Abstract
A novel biosynthetic strategy is described for the preparation of deuterated proteins containing protons at the ring carbons of Phe, Tyr and Trp, using the aromatic amino acid precursor shikimic acid. Specific protonation at aromatic side chains, with complete deuteration at C(alpha/beta) positions was achieved in proteins overexpressed in bacteria grown in shikimate-supplemented D2O medium. Co-expression of a shikimate transporter in prototrophic bacteria resulted in protonation levels of 62-79%, whereas complete labeling was accomplished using shikimate auxotrophic bacteria. Our labeling protocol permits the measurement of important aromatic side chain derived distance restraints in perdeuterated proteins that could be utilized to enhance the accuracy of NMR structures calculated using low densities of NOEs from methyl selectively protonated samples.
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