Crystal Structure of Pyrrolysyl-tRNA Synthetase from a Methanogenic Archaeon ISO4-G1 and Its Structure-Based Engineering for Highly-Productive Cell-Free Genetic Code Expansion with Non-Canonical Amino Acids.

ABSTRACT: Pairs of pyrrolysyl-tRNA synthetase (PylRS) and tRNA^Pyl from Methanosarcina mazei and Methanosarcina barkeri are widely used for site-specific incorporations of non-canonical amino acids into proteins (genetic code expansion). Previously, we achieved full productivity of cell-free protein synthesis for bulky non-canonical amino acids, including N^ε-((((E)-cyclooct-2-en-1-yl)oxy)carbonyl)-L-lysine (TCO*Lys), by using Methanomethylophilus alvus PylRS with structure-based mutations in and around the amino acid binding pocket (first-layer and second-layer mutations, respectively). Recently, the PylRS·tRNA^Pyl pair from a methanogenic archaeon ISO4-G1 was used for genetic code expansion. In the present study, we determined the cr

SUBMITTER: Yanagisawa T 

PROVIDER: S-EPMC10094482 | biostudies-literature | 2023 Mar

REPOSITORIES: biostudies-literature