Engineering mutually orthogonal PylRS/tRNA pairs for dual encoding of functional histidine analogues.

ABSTRACT: The availability of an expanded genetic code opens exciting new opportunities in enzyme design and engineering. In this regard histidine analogues have proven particularly versatile, serving as ligands to augment metalloenzyme function and as catalytic nucleophiles in designed enzymes. The ability to genetically encode multiple functional residues could greatly expand the range of chemistry accessible within enzyme active sites. Here, we develop mutually orthogonal translation components to selectively encode two structurally similar histidine analogues. Transplanting known mutations from a promiscuous Methanosarcina mazei pyrrolysyl-tRNA synthetase (MmPylRS^IFGFF ) into a single domain PylRS from Methanomethylophilus alvus (MaPylRS^IFGFF ) provided a variant with impro

SUBMITTER: Taylor CJ 

PROVIDER: S-EPMC10127257 | biostudies-literature | 2023 May

REPOSITORIES: biostudies-literature