NON-NATURAL AMINO ACID tRNA SYNTHETASES FOR PARA-METHYLAZIDO-L-PHENYLALANINE

摘要

The invention provides methods and compositions for a mutein aminoacyl-tRNA synthetase that preferentially charges a tRNA with a non-natural amino acid. Also provided are methods for incorporating the non-natural amino acid, para-methylazido-L-phenylalanine into a protein and further conjugating a biologically active adduct to the para-methylazido-L-phenylalanine.

主权项

1. A composition comprising an aminoacyl-tRNA synthetase (RS) wherein the RS:
i) preferentially aminoacylates to a degree of greater than 90% a tRNA with para-methylazido-L-phenylalanine (pAMF) compared to the 20 common naturally occurring amino acids; ii) has a sequence identity of over 80% to Methanococcus jannaschii tyrosyl tRNA synthetase (TyrRS) having SEQ NO:1; and iii) using SEQ ID NO: 1 as a reference sequence, has:
a) at position L65 amino acid: A or V;b) at position F108 amino acid: Y or W;c) at position D158 amino acid: A;d) at position Y32 amino acid: T or V or A; ande) at position I159 amino acid: S or G or V.