ArticleRNA (New York, N.Y.)2007
Improved amber and opal suppressor tRNAs for incorporation of unnatural amino acids in vivo. Part 2: evaluating suppression efficiency.
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 13 papers.
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Who cites it
13 citing papers in PubMed, 21 citations in OpenAlex.
- 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
- 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
- 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
- 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 1: minimizing misacylation.RNA (New York, N.Y.) · 2007Article
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Authors and funding
3 authors at 1 institution in 1 country.
Funding
Abstract
The incorporation of unnatural amino acids into proteins is a valuable tool for addition of biophysical probes, bio-orthogonal functionalities, and photoreactive cross-linking agents, although these approaches often require quantities of protein that are difficult to access with chemically aminoacylated tRNAs. THG73 is an amber suppressor tRNA that has been used extensively, incorporating over 100 residues in 20 proteins. In vitro studies have shown that the Escherichia coli Asn amber suppressor (ENAS) suppresses better than THG73. However, we report here that ENAS suppresses with <26% of the efficiency of THG73 in Xenopus oocytes. We then tested the newly developed Tetrahymena thermophila Gln amber suppressor (TQAS) tRNA library, which contains mutations in the second to fourth positions of the acceptor stem. The acceptor stem mutations have no adverse effect on suppression efficiency and, in fact, can increase the suppression efficiency. Combining mutations causes an averaging of suppression efficiency, and increased suppression efficiency does not correlate with increased DeltaG of the acceptor stem. We created a T. thermophila opal suppressor, TQOpS', which shows approximately 50% suppression efficiency relative to THG73. The TQAS tRNA library, composed of functional suppressor tRNAs, has been created and will allow for screening in eukaryotic cells, where rapid analysis of large libraries is not feasible.
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