Expanded ribosomal synthesis of non-standard cyclic backbones in vitro

Abstract The ribosome polymerizes L-α-amino acids into polypeptides, catalyzing peptide bond formation through aminolysis. This process is facilitated by entropy trapping within its peptidyl transferase center (PTC). In this research, we harness this capability to synthesize polymers containing cycl...

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Main Authors: Kanghun Lee, Hyeongwoo Park, Ravi Kumar Devarapalli, Dahye Im, Jongcheol Seo, Joongoo Lee
Format: Article
Language:English
Published: Nature Portfolio 2025-05-01
Series:Nature Communications
Online Access:https://doi.org/10.1038/s41467-025-60126-4
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author Kanghun Lee
Hyeongwoo Park
Ravi Kumar Devarapalli
Dahye Im
Jongcheol Seo
Joongoo Lee
author_facet Kanghun Lee
Hyeongwoo Park
Ravi Kumar Devarapalli
Dahye Im
Jongcheol Seo
Joongoo Lee
author_sort Kanghun Lee
collection DOAJ
description Abstract The ribosome polymerizes L-α-amino acids into polypeptides, catalyzing peptide bond formation through aminolysis. This process is facilitated by entropy trapping within its peptidyl transferase center (PTC). In this research, we harness this capability to synthesize polymers containing cyclic motifs in the backbone. We design 26 non-canonical monomers (ncMs) with two distinct substrates: dicarboxylic esters and hydrazinoesters, each containing bifunctional moieties that undergo ring-closing reactions through multiple aminolysis reactions. Using a cell-free system that enables the consecutive incorporation of these ncMs into a growing peptide, we discover that the ribosome can produce 5- and 6-membered cyclic backbones, which have never been reported. We also demonstrate that the formation of such cyclic backbones within the ribosome is tunable by altering the substituents of dicarboxylic esters. This discovery expands the range of non-standard backbones that can be synthesized by the ribosome and motivates future research towards expanding ribosome-mediated chemistries for biopolymer synthesis.
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institution OA Journals
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record_format Article
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spelling doaj-art-483615c096ff405ebb7fc351a668bc1b2025-08-20T02:03:31ZengNature PortfolioNature Communications2041-17232025-05-0116111410.1038/s41467-025-60126-4Expanded ribosomal synthesis of non-standard cyclic backbones in vitroKanghun Lee0Hyeongwoo Park1Ravi Kumar Devarapalli2Dahye Im3Jongcheol Seo4Joongoo Lee5Division of Interdisciplinary Bioscience and Bioengineering (I-Bio), Pohang University of Science and Technology (POSTECH)Division of Interdisciplinary Bioscience and Bioengineering (I-Bio), Pohang University of Science and Technology (POSTECH)Department of Chemical Engineering, Pohang University of Science and Technology (POSTECH)Department of Chemistry, Pohang University of Science and Technology (POSTECH)Division of Interdisciplinary Bioscience and Bioengineering (I-Bio), Pohang University of Science and Technology (POSTECH)Division of Interdisciplinary Bioscience and Bioengineering (I-Bio), Pohang University of Science and Technology (POSTECH)Abstract The ribosome polymerizes L-α-amino acids into polypeptides, catalyzing peptide bond formation through aminolysis. This process is facilitated by entropy trapping within its peptidyl transferase center (PTC). In this research, we harness this capability to synthesize polymers containing cyclic motifs in the backbone. We design 26 non-canonical monomers (ncMs) with two distinct substrates: dicarboxylic esters and hydrazinoesters, each containing bifunctional moieties that undergo ring-closing reactions through multiple aminolysis reactions. Using a cell-free system that enables the consecutive incorporation of these ncMs into a growing peptide, we discover that the ribosome can produce 5- and 6-membered cyclic backbones, which have never been reported. We also demonstrate that the formation of such cyclic backbones within the ribosome is tunable by altering the substituents of dicarboxylic esters. This discovery expands the range of non-standard backbones that can be synthesized by the ribosome and motivates future research towards expanding ribosome-mediated chemistries for biopolymer synthesis.https://doi.org/10.1038/s41467-025-60126-4
spellingShingle Kanghun Lee
Hyeongwoo Park
Ravi Kumar Devarapalli
Dahye Im
Jongcheol Seo
Joongoo Lee
Expanded ribosomal synthesis of non-standard cyclic backbones in vitro
Nature Communications
title Expanded ribosomal synthesis of non-standard cyclic backbones in vitro
title_full Expanded ribosomal synthesis of non-standard cyclic backbones in vitro
title_fullStr Expanded ribosomal synthesis of non-standard cyclic backbones in vitro
title_full_unstemmed Expanded ribosomal synthesis of non-standard cyclic backbones in vitro
title_short Expanded ribosomal synthesis of non-standard cyclic backbones in vitro
title_sort expanded ribosomal synthesis of non standard cyclic backbones in vitro
url https://doi.org/10.1038/s41467-025-60126-4
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AT dahyeim expandedribosomalsynthesisofnonstandardcyclicbackbonesinvitro
AT jongcheolseo expandedribosomalsynthesisofnonstandardcyclicbackbonesinvitro
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