Resolving the impasse in expanding genetic codes
In the realm of genetic code expansion, a groundbreaking study has shattered the constraints limiting the synthesis of proteins to only α-amino acids. The traditional approach to incorporating non-canonical monomers (ncMs) into proteins has been hindered by the inability to expand beyond α-amino acids due to the intricate dance between aminoacyl-tRNA synthetases (aaRS) and the ribosome during protein synthesis.
However, in a recent publication in Nature, Dunkelmann and colleagues unveiled a novel method dubbed 'tRNA display' that broke the deadlock. By decoupling the evolution of aaRS from ribosome-dependent selection, they devised a strategy to engineer Escherichia coli's translation apparatus to incorporate β-linked and α,α-disubstituted monomers into ribosome-synthesized proteins.
The process involves acylating an ncM onto a tRNA that reads the amber codon UAG, which typically signifies termination of protein synthesis. The team leveraged the PylRS-tRNA system, orthogonal to E. coli, and utilized tRNA display to evolve aaRS variants capable of acylating specific ncMs onto tRNA without the ribosome readout. This innovative approach led to the successful incorporation of diverse backbone-modified ncMs into proteins, including β-amino acids and α,α-disubstituted amino acids, with impressive efficiency.
The implications of this breakthrough are profound. The ability to incorporate a broader range of monomers into proteins opens up new possibilities for the development of protein-based medicines and biocatalysts with unique properties. Proteins containing non-canonical monomers could lead to therapeutics with reduced immunogenicity and increased stability, as well as novel industrial biocatalysts with enhanced regio- and stereochemistry.
Moving forward, further advancements in engineered ribosomes that can accommodate backbone-modified amino acids will be crucial. By dissecting the underlying mechanisms and structural features that enable the ribosome to synthesize proteins with diverse monomers, researchers aim to optimize the activity of these engineered ribosomes and pave the way for a new era in protein engineering.
The study by Dunkelmann et al. marks a pivotal moment in genetic code expansion, propelling us towards a future where the possibilities of protein synthesis are boundless. With the shackles of traditional constraints broken, the horizon of bioengineering shines brighter than ever before.
Source: https://www.nature.com/articles/s41589-024-01579-4
However, in a recent publication in Nature, Dunkelmann and colleagues unveiled a novel method dubbed 'tRNA display' that broke the deadlock. By decoupling the evolution of aaRS from ribosome-dependent selection, they devised a strategy to engineer Escherichia coli's translation apparatus to incorporate β-linked and α,α-disubstituted monomers into ribosome-synthesized proteins.
The process involves acylating an ncM onto a tRNA that reads the amber codon UAG, which typically signifies termination of protein synthesis. The team leveraged the PylRS-tRNA system, orthogonal to E. coli, and utilized tRNA display to evolve aaRS variants capable of acylating specific ncMs onto tRNA without the ribosome readout. This innovative approach led to the successful incorporation of diverse backbone-modified ncMs into proteins, including β-amino acids and α,α-disubstituted amino acids, with impressive efficiency.
The implications of this breakthrough are profound. The ability to incorporate a broader range of monomers into proteins opens up new possibilities for the development of protein-based medicines and biocatalysts with unique properties. Proteins containing non-canonical monomers could lead to therapeutics with reduced immunogenicity and increased stability, as well as novel industrial biocatalysts with enhanced regio- and stereochemistry.
Moving forward, further advancements in engineered ribosomes that can accommodate backbone-modified amino acids will be crucial. By dissecting the underlying mechanisms and structural features that enable the ribosome to synthesize proteins with diverse monomers, researchers aim to optimize the activity of these engineered ribosomes and pave the way for a new era in protein engineering.
The study by Dunkelmann et al. marks a pivotal moment in genetic code expansion, propelling us towards a future where the possibilities of protein synthesis are boundless. With the shackles of traditional constraints broken, the horizon of bioengineering shines brighter than ever before.
Source: https://www.nature.com/articles/s41589-024-01579-4
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