An Evolved Methanomethylophilus alvus Pyrrolysyl-tRNA Synthetase/tRNA Pair Is Highly Active and Orthogonal in Mammalian Cells.

ABSTRACT: We recently characterized a new class of pyrrolysyl-tRNA synthetase (PylRS)/^PyltRNA pairs from Methanomassiliicocales that are active and orthogonal in Escherichia coli. The aminoacyl-tRNA synthetases (aaRSs) of these pairs lack the N-terminal domain that is essential for tRNA recognition and in vivo activity in the Methanosarcina mazei ( Mm) PylRS but share a homologous active site with MmPylRS; this facilitates the transplantation of mutations discovered with existing PylRS systems into the new PylRS systems to reprogram their substrate specificity for the incorporation of noncanonical amino acids (ncAAs). Several of the new PylRS/^PyltRNA pairs, or their evolved variants [including Methanomethylophilus alvus ( Ma) PylRS/ Ma^PyltRNA(6)_CUA], are m

SUBMITTER: Beranek V 

PROVIDER: S-EPMC6365905 | biostudies-literature | 2019 Feb

REPOSITORIES: biostudies-literature