NON-NATURAL AMINO ACID tRNA SYNTHETASES FOR PYRIDYL TETRAZINE

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 acids, pyridinyl-amino tetrazine amino acids, into a protein, and further conjugating a biologically active adduct to the pyridinyl-amino tetrazine.

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 (S)-2-amino-3-(5-((6-methyl-1,2,4,5-tetrazin-3-ylamino)methyl)pyridin-2-yl)propanoic acid or a compound having the formula: 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 ID NO: 1; and iii) using SEQ ID NO: 1 as a reference sequence, has:
a) at position Y32 amino acid: L;b) at position L65 amino acid: V;c) at position H70 amino acid: Ad) at position F108 amino acid: W;e) at position Q109 amino acid: S;f) at position D158 amino acid: V;g) at position 1159 amino acid: A; andh) at position L162 amino acid: S or V.