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-Structure paper
| タイトル | Co-Translational Incorporation of - and -β-Hydroxy Acids : A Structural and Biochemical Study on the Ribosome. |
|---|---|
| ジャーナル・号・ページ | J Am Chem Soc, Vol. 148, Issue 9, Page 9393-9399, Year 2026 |
| 掲載日 | 2026年3月11日 |
著者 | Chandrima Majumdar / Alexandra D Kent / Noah X Hamlish / Cathy Zhu / Katelyn A Fitzgerald / Jamie H D Cate / Alanna Schepartz / ![]() |
| PubMed 要旨 | Engineering the translation apparatus to accept backbone-modified amino acid analogues would enable the programmed synthesis of sequence-defined biopolymers with tunable properties. β-Hydroxy acids ...Engineering the translation apparatus to accept backbone-modified amino acid analogues would enable the programmed synthesis of sequence-defined biopolymers with tunable properties. β-Hydroxy acids are of particular interest because they could support the programmed biosynthesis of both biocompatible polyester materials as well as natural product-like depsipeptides. Previous work has reported that both enantiomers of β-hydroxy-N-Boc-lysine (β-OH-BocK) are substrates for the orthogonal pyrrolysyl-tRNA synthetase (PylRS)/tRNA pair, but only one enantiomer is introduced into protein . Here we make use of high-resolution cryogenic electron microscopy (cryo-EM) to determine the structural basis for this observation. These structures reveal both β-OH-BocK isomers equally well-positioned within the ribosomal A site regardless of stereochemistry. Consistent with this observation, translation reactions charged with tRNAs acylated with - or -β-OH-BocK produced roughly equal amounts of translated product when quantified on the basis of either mass spectrometry or luminescence. Together, these experiments imply that the substantial preferential incorporation of one enantiomer over the other observed previously results primarily from deficiencies in the steps that precede bond formation by the ribosome. Indeed, as predicted by this work and demonstrated in an accompanying paper (Soni, C. Co-Translational Incorporation of ()- and ()-β-Hydroxyacids : Directed Evolution of Efficient Aminoacyl-tRNA Synthetases. 2026, 148, 10.1021/jacs.5c18595), when cells are provided with an active and orthogonal aminoacyl-tRNA synthetase/tRNA pair that accepts both - and -β-OH-BocK as substrates, both monomers are introduced into protein in good yield and with high fidelity. |
リンク | J Am Chem Soc / PubMed:41757712 / PubMed Central |
| 手法 | EM (単粒子) |
| 解像度 | 2.14 - 2.56 Å |
| 構造データ | EMDB-70050, PDB-9o2x: EMDB-70051, PDB-9o2y: ![]() EMDB-70052: 50S focus refined map of E.coli 70S ribosome complexed with P-site fMet-tRNAfMet and A-site S-beta(2)hydroxyBocK-tRNAPyl ![]() EMDB-70054: 30S focus refined map of E.coli 70S ribosome complexed with P-site fMet-tRNAfMet and A-site S-beta(2)hydroxyBocK-tRNAPyl ![]() EMDB-70055: 50S focus refined map of E.coli 70S ribosome complexed with P-site fMet-tRNAfMet and A-site R-beta(2)hydroxyBocK-tRNAPyl ![]() EMDB-70056: 30S focus refined map of E.coli 70S ribosome complexed with P-site fMet-tRNAfMet and A-site R-beta(2)hydroxyBocK-tRNAPyl ![]() EMDB-70477: 70S global refined map of E.coli 70S ribosome complexed with P-site fMet-tRNAfMet and A-site S-beta(2)hydroxyBocK-tRNAPyl ![]() EMDB-70478: 70S global refined map of E.coli 70S ribosome complexed with P-site fMet-tRNAfMet and A-site R-beta(2)hydroxyBocK-tRNAPyl |
| 化合物 | ![]() ChemComp-ZN: ![]() ChemComp-MG: ![]() ChemComp-FME: ![]() ChemComp-K: ![]() ChemComp-PAR: ![]() ChemComp-SPD: ![]() ChemComp-SPM: ![]() PDB-1b71: ![]() ChemComp-HOH: ![]() PDB-1b70: |
| 由来 |
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キーワード | RIBOSOME / 70S / unnatural amino acids / non-natural monomers / beta-2-hydroxy acids / unnatural monomers |
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methanomethylophilus alvi (古細菌)
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