Efficient Incorporation of DOPA into Proteins Free from Competition with Endogenous Translation Termination Machinery.

ABSTRACT: 3,4-Dihydroxy-L-phenylalanine (DOPA) is a promising noncanonical amino acid (ncAA) that introduces novel catechol chemical features into proteins, expanding their functional potential. However, the most common approach to incorporating ncAAs into proteins relies on stop codon suppression, which is often limited by the competition of endogenous translational termination machinery. Here, we employed a special in vitro protein expression system that facilitates the efficiency of DOPA incorporation into proteins by removing essential Class I peptide release factors through targeted degradation. In the absence of both RF1 and RF2, we successfully demonstrated DOPA incorporation at all three stop codons (TAG, TAA, and TGA). By optimizing the concentration of engineered DOPA-specific aminoacyl-tR

SUBMITTER: Yang Y 

PROVIDER: S-EPMC11939889 | biostudies-literature | 2025 Mar

REPOSITORIES: biostudies-literature