Incorporation of unnatural amino acids having an orthogonal functional group into polypeptides using orthogonal tRNA/tRNA synthetase pairs

摘要

A method of making a polypeptide comprising an orthogonal functional group, said orthogonal functional group comprising an aliphatic amino acid; the method comprising providing a host cell; providing a nucleic acid encoding the polypeptide of interest; providing a tRNA-tRNA synthetase pair orthogonal to said host cell; adding an amino acid or amino acid derivative comprising the orthogonal functional group of interest (e.g. alkyne, azide or ketone group), wherein said amino acid or amino acid derivative has an aliphatic carbon backbone; and incubating to allow incorporation of said amino acid or amino acid derivative into the polypeptide of interest via the orthogonal tRNA-tRNA synthetase pair. Also claimed are amino acids comprising an aliphatic functional group; a polypeptide comprising N6-[(2-propynyloxy)carbonyl]-L-lysine, N6-[(2-azidoethoxy)carbonyl]-L-lysine or (S)-2-amino-6-((pent-4-enyloxy)carbonylamino)hexanoic acid; and tRNA synthetase polypeptides and nucleic acids.