Engineering the Ribosomal Translation System to Introduce Non‐proteinogenic Amino Acids into Peptides

Katoh, Takayuki

doi:10.1002/9783527823987.vol2_c5

Cited by 1 publication

(1 citation statement)

References 83 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…As natural aminoacyl-tRNA synthetases (ARS) cannot efficiently charge nPAAs onto tRNAs, alternative methods for preparing nPAA-tRNAs are necessary. Accordingly, the following methodologies have been developed so far: (1) the use of an aminoacylation ribozyme named flexizyme, which is used in the studies by Katoh et al for the preparation of d -α- and β-aminoacyl-tRNAs [23,24,29,30,31], (2) charging nPAAs by means of mutant ARSs (used in the studies by Lee et al and Fan et al) [13,14,32,33,34,35,36], (3) the ligation of nPAA-pdCpA dinucleotides with tRNA lacking the 3′-terminal CA dinucleotide (used in the studies by Doi et al, Dedkova et al, and Maini et al) [12,19,37,38,39], and (4) the post-aminoacylation modification of PAA-tRNA (used in the study by Haruna et al) [1,17,40,41,42,43]. However, the details of these methodologies are discussed elsewhere [32].…”

Section: Perspectivesmentioning

confidence: 99%

Engineering Translation Components Improve Incorporation of Exotic Amino Acids

Katoh¹,

Suga²

2019

IJMS

Self Cite

View full text Add to dashboard Cite

Methods of genetic code manipulation, such as nonsense codon suppression and genetic code reprogramming, have enabled the incorporation of various nonproteinogenic amino acids into the peptide nascent chain. However, the incorporation efficiency of such amino acids largely varies depending on their structural characteristics. For instance, l-α-amino acids with artificial, bulky side chains are poorer substrates for ribosomal incorporation into the nascent peptide chain, mainly owing to the lower affinity of their aminoacyl-tRNA toward elongation factor-thermo unstable (EF-Tu). Phosphorylated Ser and Tyr are also poorer substrates for the same reason; engineering EF-Tu has turned out to be effective in improving their incorporation efficiencies. On the other hand, exotic amino acids such as d-amino acids and β-amino acids are even poorer substrates owing to their low affinity to EF-Tu and poor compatibility to the ribosome active site. Moreover, their consecutive incorporation is extremely difficult. To solve these problems, the engineering of ribosomes and tRNAs has been executed, leading to successful but limited improvement of their incorporation efficiency. In this review, we comprehensively summarize recent attempts to engineer the translation systems, resulting in a significant improvement of the incorporation of exotic amino acids.

show abstract