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  A kinetic safety gate controlling the delivery of unnatural amino acids to the ribosome.

Mittelstät, J., Konevega, A. L., & Rodnina, M. V. (2013). A kinetic safety gate controlling the delivery of unnatural amino acids to the ribosome. Journal of the American Chemical Society, 135(45), 17031-17038. doi:10.1021/ja407511q.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0014-6239-6 Version Permalink: http://hdl.handle.net/11858/00-001M-0000-0028-3B08-7
Genre: Journal Article

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http://pubs.acs.org/doi/pdf/10.1021/ja407511q (Publisher version)
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Mittelstät, J.1, Author              
Konevega, A. L.1, Author              
Rodnina, M. V.1, Author              
Affiliations:
1Department of Physical Biochemistry, MPI for biophysical chemistry, Max Planck Society, ou_578598              

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 Abstract: Improving the yield of unnatural amino acid incorporation is an important challenge in producing novel designer proteins with unique chemical properties. Here we examine the mechanisms that restrict the incorporation of the fluorescent unnatural amino acid εNH2-Bodipy576/589-lysine (BOP-Lys) into a model protein. While the delivery of BOP-Lys-tRNALys to the ribosome is limited by its poor binding to elongation factor Tu (EF-Tu), the yield of incorporation into peptide is additionally controlled at the step of BOP-Lys-tRNA release from EF-Tu into the ribo-some. The unnatural amino acid appears to disrupt the interactions that balance the strength of tRNA binding to EF-Tu–GTP with the velocity of tRNA dissociation from EF-Tu–GDP on the ribosome, which ensure uniform incorporation of standard amino acids. Circumventing this potential quality control checkpoint that specifically prevents incorporation of unnatural amino acids into proteins may provide a new strategy to increase yields of unnatural polymers.

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Language(s): eng - English
 Dates: 2013-09-30
 Publication Status: Published online
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 Rev. Method: Peer
 Identifiers: DOI: 10.1021/ja407511q
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Title: Journal of the American Chemical Society
Source Genre: Journal
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Pages: - Volume / Issue: 135 (45) Sequence Number: - Start / End Page: 17031 - 17038 Identifier: -