Site‐specific incorporation of photofunctional nonnatural amino acids into a polypeptide through in vitro protein biosynthesis

Abstract
Nonnatural amino acids with photofunctional groups were incorporated site‐specifically into a polypeptide by using in vitro protein synthesizing system. The nonnatural amino acids were attached to tRNACCU through chemical misacylation method, and added to the in vitro system with a mRNA containing a single AGG codon. l‐p‐Phenylazophenylalanine, l‐2‐anthrylalanine, l‐1‐naphthylalanine, l‐2‐naphthylalanine and l‐p‐biphenylalanine were successfully incorporated into a polypeptide, but l‐1‐pyrenylalanine was not. The polypeptides containing the nonnatural amino acids showed photofunctionalities.