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n recent years, many laboratories have employed ribosomemediated protein synthesis to produce proteins containing
non-proteinogenic amino acids, enabling a more detailed study
of protein structure, function, and dynamics.1 While a broad
range of amino acids can now be incorporated into proteins,
the incorporation of -amino acids containing large and
negatively charged side chains is still challenging,2 as is protein
biosynthesis with non -amino acids.3
Recently, we have described the facilitated incorporation of
non-proteinogenic amino acids by the use of modied
ribosomes in which key regions of the Escherichia coli 23S
2015 American Chemical Society
DOI: 10.1021/jacs.5b03135
J. Am. Chem. Soc. 2015, 137, 1120611209
Communication
DOI: 10.1021/jacs.5b03135
J. Am. Chem. Soc. 2015, 137, 1120611209
Communication
region 1
region 2
(20572063)
(25022507)
amino acid
5
UGCGUGG
AGUGAGA
ACGAAG
AUCCGA
0.8 0.2
2.2 1.0
15.3 2.5
14.0 3.0
DOI: 10.1021/jacs.5b03135
J. Am. Chem. Soc. 2015, 137, 1120611209
Communication
ex/abs,
nm
quantum yield, em
GFPwt
GFP66oxazole5
GFP66Gly39oxazole5
395
310
310
25,000
90,300
50,200
ASSOCIATED CONTENT
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AUTHOR INFORMATION
Corresponding Author
*sidhecht@asu.edu
Notes
ACKNOWLEDGMENTS
This work was supported by National Institutes of Health
Research Grant GM103861, awarded by the National Institute
of General Medical Sciences. We thank Dr. Sriloy Dey for
assistance in preparing a synthetic intermediate.
REFERENCES
DOI: 10.1021/jacs.5b03135
J. Am. Chem. Soc. 2015, 137, 1120611209