Incorporation of Two Different Nonnatural Amino Acids Independently into a Single Protein through Extension of the Genetic Code

1999 ◽  
Vol 121 (51) ◽  
pp. 12194-12195 ◽  
Author(s):  
Takahiro Hohsaka ◽  
Yuki Ashizuka ◽  
Hiroshi Sasaki ◽  
Hiroshi Murakami ◽  
Masahiko Sisido
Keyword(s):  
Amino Acids ◽  
Genetic Code ◽  
Nonnatural Amino Acids ◽  
Single Protein

scholarly journals Transferability of N-terminal mutations of pyrrolysyl-tRNA synthetase in one species to that in another species on unnatural amino acid incorporation efficiency

Amino Acids ◽  
2020 ◽  
Author(s):  
Thomas L. Williams ◽  
Debra J. Iskandar ◽  
Alexander R. Nödling ◽  
Yurong Tan ◽  
Louis Y. P. Luk ◽  
...  
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Genetic Code ◽  
Unnatural Amino Acids ◽  
Trna Synthetase ◽  
Unnatural Amino Acid ◽  
Amino Acid Incorporation ◽  
Acid Incorporation ◽  
Genetic Code Expansion ◽  
Incorporation Efficiency

AbstractGenetic code expansion is a powerful technique for site-specific incorporation of an unnatural amino acid into a protein of interest. This technique relies on an orthogonal aminoacyl-tRNA synthetase/tRNA pair and has enabled incorporation of over 100 different unnatural amino acids into ribosomally synthesized proteins in cells. Pyrrolysyl-tRNA synthetase (PylRS) and its cognate tRNA from Methanosarcina species are arguably the most widely used orthogonal pair. Here, we investigated whether beneficial effect in unnatural amino acid incorporation caused by N-terminal mutations in PylRS of one species is transferable to PylRS of another species. It was shown that conserved mutations on the N-terminal domain of MmPylRS improved the unnatural amino acid incorporation efficiency up to five folds. As MbPylRS shares high sequence identity to MmPylRS, and the two homologs are often used interchangeably, we examined incorporation of five unnatural amino acids by four MbPylRS variants at two temperatures. Our results indicate that the beneficial N-terminal mutations in MmPylRS did not improve unnatural amino acid incorporation efficiency by MbPylRS. Knowledge from this work contributes to our understanding of PylRS homologs which are needed to improve the technique of genetic code expansion in the future.

On the Optimization of the Physicochemical Distances between Amino Acids in the Evolution of the Genetic Code

1994 ◽  
Vol 168 (1) ◽  
pp. 43-51 ◽  
Author(s):  
Massimo Di Giulio ◽  
M.Rosaria Capobianco ◽  
Mario Medugno
Keyword(s):  
Amino Acids ◽  
Genetic Code

scholarly journals Expanding the genetic code of Salmonella with non-canonical amino acids

2016 ◽  
Vol 6 (1) ◽  
Author(s):  
Qinglei Gan ◽  
Brent P. Lehman ◽  
Thomas A. Bobik ◽  
Chenguang Fan
Keyword(s):  
Amino Acids ◽  
Genetic Code
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