Orthogonal Protein Translation Using Pyrrolysyl-tRNA Synthetases for Single- and Multiple-Noncanonical Amino Acid Mutagenesis

2016 ◽  
pp. 1-19 ◽  
Author(s):  
Tobias Baumann ◽  
Matthias Exner ◽  
Nediljko Budisa
Keyword(s):  
Amino Acid ◽  
Protein Translation ◽  
Trna Synthetases ◽  
Amino Acid Mutagenesis ◽  
Noncanonical Amino Acid

scholarly journals Expanding the Scope of Single- and Double-Noncanonical Amino Acid Mutagenesis in Mammalian Cells Using Orthogonal Polyspecific Leucyl-tRNA Synthetases

Biochemistry ◽  
2017 ◽  
Vol 57 (4) ◽  
pp. 441-445 ◽  
Author(s):  
Yunan Zheng ◽  
Raja Mukherjee ◽  
Melissa A. Chin ◽  
Peter Igo ◽  
Martin J. Gilgenast ◽  
...  
Keyword(s):  
Amino Acid ◽  
Mammalian Cells ◽  
Trna Synthetases ◽  
Amino Acid Mutagenesis ◽  
Noncanonical Amino Acid

scholarly journals Alteration in H-bond strength affects the stability of codon-anticodon interaction at in-frame UAG stop codon during in vitro translation

2021 ◽  
Author(s):  
Purnima Mala ◽  
Ishu Saraogi
Keyword(s):  
Amino Acid ◽  
Stop Codon ◽  
Protein Translation ◽  
In Vitro Translation ◽  
Natural Amino Acid ◽  
Additional Hydrogen Bond ◽  
Amino Acid Mutagenesis ◽  
The Stability ◽  
Decoding Ability

We have studied the decoding ability of a non-standard nucleobase modified tRNA for non-natural amino acid mutagenesis. The insertion of 2, 6-diaminopurine (D) base at the 3rd position of a tRNA anticodon enabled us to evaluate the effect of an additional hydrogen bond during translation. The presence of D at the tRNA anticodon led to stabilization of the codon-anticodon interaction due to an additional H-bond between the N2-exocyclic amine of D and the C2 carbonyl group of uracil during protein translation. While decoding UAG codons using stop codon suppression methodology, the enhanced codon-anticodon interaction improved codon readthrough and synthesis of modified protein with a non-natural amino acid at multiple sites. Our findings imply that the number of hydrogen bonds at the tRNA-mRNA duplex interface is an important criterion during mRNA decoding and improves protein translation at multiple UAG stop sites. This work provides valuable inputs towards improved non-natural amino acid mutagenesis for creating functional proteins.

Manipulating Unconventional CH-Based Hydrogen Bonding in a Methyltransferase via Noncanonical Amino Acid Mutagenesis

2014 ◽  
Vol 9 (8) ◽  
pp. 1692-1697 ◽  
Author(s):  
Scott Horowitz ◽  
Upendra Adhikari ◽  
Lynnette M. A. Dirk ◽  
Paul A. Del Rizzo ◽  
Ryan A. Mehl ◽  
...  
Keyword(s):  
Hydrogen Bonding ◽  
Amino Acid ◽  
Amino Acid Mutagenesis ◽  
Noncanonical Amino Acid

scholarly journals Defining the current scope and limitations of dual noncanonical amino acid mutagenesis in mammalian cells

2017 ◽  
Vol 8 (10) ◽  
pp. 7211-7217 ◽  
Author(s):  
Yunan Zheng ◽  
Partha Sarathi Addy ◽  
Raja Mukherjee ◽  
Abhishek Chatterjee
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Mammalian Cells ◽  
Noncanonical Amino Acids ◽  
Amino Acid Mutagenesis ◽  
Noncanonical Amino Acid

We systematically evaluate potential platforms for site-specifically incorporating two distinct noncanonical amino acids into proteins expressed in mammalian cells with optimal fidelity and efficiency – a technology that will have many enabling applications.

scholarly journals Aminoacyl tRNA Synthetases as Malarial Drug Targets: A Comparative Bioinformatics Study

2018 ◽  
Author(s):  
Dorothy Wavinya Nyamai ◽  
Özlem Tastan Bishop
Keyword(s):  
Amino Acid ◽  
Drug Targets ◽  
Motif Discovery ◽  
Protein Translation ◽  
Trna Synthetase ◽  
Potential Drug ◽  
Multiple Sequence ◽  
Trna Synthetases ◽  
Aminoacyl Trna Synthetases ◽  
Potential Drug Targets

AbstractTreatment of parasitic diseases has been challenging due to the development of drug resistance by parasites, and thus there is need to identify new class of drugs and drug targets. Protein translation is important for survival of plasmodium and the pathway is present in all the life cycle stages of the plasmodium parasite. Aminoacyl tRNA synthetases are primary enzymes in protein translation as they catalyse the first reaction where an amino acid is added to the cognate tRNA. Currently, there is limited research on comparative studies of aminoacyl tRNA synthetases as potential drug targets. The aim of this study is to understand differences between plasmodium and human aminoacyl tRNA synthetases through bioinformatics analysis. Plasmodium falciparum, P. fragile, P. vivax, P. ovale, P. knowlesi, P. bergei, P. malariae and human aminoacyl tRNA synthetase sequences were retrieved from UniProt database and grouped into 20 families based on amino acid specificity. Despite functional and structural conservation, multiple sequence analysis, motif discovery, pairwise sequence identity calculations and molecular phylogenetic analysis showed striking differences between parasite and human proteins. Prediction of alternate binding sites revealed potential druggable sites in PfArgRS, PfMetRS and PfProRS at regions that were weakly conserved when compared to the human homologues. These differences provide a basis for further exploration of plasmodium aminoacyl tRNA synthetases as potential drug targets.

Performance of optimized noncanonical amino acid mutagenesis systems in the absence of release factor 1

2016 ◽  
Vol 12 (6) ◽  
pp. 1746-1749 ◽  
Author(s):  
Yunan Zheng ◽  
Marc J. Lajoie ◽  
James S. Italia ◽  
Melissa A. Chin ◽  
George M. Church ◽  
...  
Keyword(s):  
Amino Acid ◽  
Release Factor ◽  
Amino Acid Incorporation ◽  
E Coli ◽  
Acid Incorporation ◽  
Amino Acid Mutagenesis ◽  
Noncanonical Amino Acid

Unconditional deletion of RF1 in a genomically recoded E. coli enables multisite noncanonical amino acid incorporation by UAG suppression.

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