“Chemical aminoacylation” of tRNA's

Protein synthesis: Twenty three amino acids and counting

“…It was recognized early on that protein synthesis is at the interface of chemistry and biology, and attempts to deviate from the constraints of the genetic code in vitro began with the combined chemical/enzymatic generation of tRNAs acylated with desired amino acid derivatives [10]. The idea that nonsense codons are ideal signals for amino acid misincorporation mediated by suppressor tRNAs is well established [11].…”

Genetic Code Expansion: A Brief History and Perspective

“…In 1978, Hecht and co-workers established a general procedure for the chemoenzymatic aminoacylation of tRNAs . T4 RNA ligase transfers an aminoacyladenylate moiety from N-blocked (with o -nitrophenylsulfenyl) aminoacylated P 1 , P 2 -bis­(5′-adenosyl)­diphosphates to tRNAs lacking the 3′-terminal adenosine.…”

Trapping Transient Protein Species by Genetic Code Expansion

“…In this approach, as for the production of natural selenocysteine-and pyrrolysine-containing proteins, the desired building block has to be coupled to a suppressor tRNA that recognizes the target stop codon. This can be accomplished either by chemical synthesis [22] or by using engineered ribozymes [23] or aminoacyl-tRNA synthetases [7,8] that catalyze the aminoacylation reaction. For in vivo experiments, suppressor tRNA/aminoacyl-tRNA synthetase (aaRS) pairs have been evolved for ncAAs that are orthogonal to the translation machinery of the respective host organism.…”