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Science 14 April 1989:
Vol. 244. no. 4901, pp. 182 - 188
DOI: 10.1126/science.2649980
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Articles

Science, Vol 244, Issue 4901, 182-188
Copyright © 1989 by American Association for the Advancement of Science

articles

A general method for site-specific incorporation of unnatural amino acids into proteins

CJ Noren, SJ Anthony-Cahill, MC Griffith, and PG Schultz

Department of Chemistry, University of California, Berkeley 94720.

A new method has been developed that makes it possible to site-specifically incorporate unnatural amino acids into proteins. Synthetic amino acids were incorporated into the enzyme beta-lactamase by the use of a chemically acylated suppressor transfer RNA that inserted the amino acid in response to a stop codon substituted for the codon encoding residue of interest. Peptide mapping localized the inserted amino acid to a single peptide, and enough enzyme could be generated for purification to homogeneity. The catalytic properties of several mutants at the conserved Phe66 were characterized. The ability to selectively replace amino acids in a protein with a wide variety of structural and electronic variants should provide a more detailed understanding of protein structure and function.


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