ArticleRNA (New York, N.Y.)2007
Improved amber and opal suppressor tRNAs for incorporation of unnatural amino acids in vivo. Part 1: minimizing misacylation.
Article in RNA (New York, N.Y.), 2007. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 20 papers.
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Who cites it
20 citing papers in PubMed, 31 citations in OpenAlex.
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- tRNA engineering strategies for genetic code expansion.Frontiers in genetics · 2024Review
- Tinker, Tailor, Soldier, Spy: The Diverse Roles That Fluorine Can Play within Amino Acid Side Chains.Molecules (Basel, Switzerland) · 2023Review
- Suppressor Mutants: History and Today's Applications.EcoSal Plus · 2021Review
- HIFs: New arginine mimic inhibitors of the Hv1 channel with improved VSD-ligand interactions.The Journal of general physiology · 2021Article
- A self-labeling protein based on the small ultra-red fluorescent protein, smURFP.RSC chemical biology · 2021Article
- Orthogonality of Pyrrolysine tRNA in the Xenopus oocyte.Scientific reports · 2018Article
- Incorporation of Non-Canonical Amino Acids.Advances in experimental medicine and biology · 2015Review
- In vivo incorporation of non-canonical amino acids by using the chemical aminoacylation strategy: a broadly applicable mechanistic tool.Chembiochem : a European journal of chemical biology · 2014Review
- The evolution of the ribosome and the genetic code.Life (Basel, Switzerland) · 2014Article
- An unusual pattern of ligand-receptor interactions for the α7 nicotinic acetylcholine receptor, with implications for the binding of varenicline.Molecular pharmacology · 2013Article
- Preparation of translationally competent tRNA by direct chemical acylation.Organic letters · 2010Article
- In vitro incorporation of nonnatural amino acids into protein using tRNA(Cys)-derived opal, ochre, and amber suppressor tRNAs.RNA (New York, N.Y.) · 2010Article
- Nicotinic pharmacophore: the pyridine N of nicotine and carbonyl of acetylcholine hydrogen bond across a subunit interface to a backbone NH.Proceedings of the National Academy of Sciences of the United States of America · 2010Article
- Chemical scale studies of the Phe-Pro conserved motif in the cys loop of Cys loop receptors.The Journal of biological chemistry · 2010Article
- Probing the role of the cation-pi interaction in the binding sites of GPCRs using unnatural amino acids.Proceedings of the National Academy of Sciences of the United States of America · 2009Article
- High throughput electrophysiology with Xenopus oocytes.Combinatorial chemistry & high throughput screening · 2009Review
- Single-molecule imaging of a fluorescent unnatural amino acid incorporated into nicotinic receptors.Biophysical journal · 2009Article
- Site-specific incorporation of unnatural amino acids into urate oxidase in Escherichia coli.Protein science : a publication of the Protein Society · 2008Article
- Improved amber and opal suppressor tRNAs for incorporation of unnatural amino acids in vivo. Part 2: evaluating suppression efficiency.RNA (New York, N.Y.) · 2007Article
Corrections and comments
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Authors and funding
3 authors at 1 institution in 1 country.
Funding
Abstract
The incorporation of unnatural amino acids site-specifically is a valuable technique for structure-function studies, incorporation of biophysical probes, and determining protein-protein interactions. THG73 is an amber suppressor tRNA used extensively for the incorporation of >100 different residues in over 20 proteins, but under certain conditions THG73 is aminoacylated in vivo by endogenous aminoacyl-tRNA synthetase. Similar aminoacylation is seen with the Escherichia coli Asn amber suppressor tRNA, which has also been used to incorporate UAAs in many studies. We now find that the natural amino acid placed on THG73 is Gln. Since the E. coli GlnRS recognizes positions in the acceptor stem, we made several acceptor stem mutations in the second to fourth positions on THG73. All mutations reduce aminoacylation in vivo and allow for the selection of highly orthogonal tRNAs. To show the generality of these mutations, we created opal suppressor tRNAs that show less aminoacylation in Xenopus oocytes relative to THG73. We have created a library of Tetrahymena thermophila Gln amber suppressor tRNAs that will be useful for determining optimal suppressor tRNAs for use in other eukaryotic cells.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.