A general and efficient route for chemical aminoacylation of transfer RNAs

Article abstract of DOI:10.1021/ja00007a055

A general and expedient procedure for the synthesis of aminoaeyl tRNA has been developed. The preparation of aminoacyl dCpA in one step and in high yield by reaction of the cyanomethyl active esters of N-protected α-amino with pdCpA is detailed. The preparation and photodeprotection of aminoacyl pdCpA derivatives containing nitroveratryl (NVOC) N-protected amino acids is also studied. The utility of the above methods for preparing aminoacyl tRNA was confirmed by enzymatically ligating NVOC-phenylalanyl pdCpA to tRNA-C_OH followed by photolysis to provide unprotected phenylalanyl tRNA. The phenylalanyl tRNA shown to be competent in an in vitro protein biosynthesis system. These protocols greatly simplify the use of chemically misacylated tRNAs in the synthesis of proteins containing unnatural ainino acids, as well as in studies of protein biosynthesis.