Ribosome-mediated biosynthesis of pyridazinone oligomers in vitro.
Ribosome-mediated biosynthesis of pyridazinone oligomers in vitro.
复制标题
DOI：
10.1038/s41467-022-33701-2
复制
发表时间：
2022-10-24
影响因子：
16.6
通讯作者：
中科院分区：
文献类型：
--
作者：
核糖体是一种大分子机器，可催化 L-α-氨基酸按序列聚合成多肽。氨基酸底物之间肽键形成的催化基于熵捕获，其中 P 位点中的转移 RNA (tRNA) 偶联酰基键和 A 位点中的 α-氨基邻接对齐底物以进行偶联。这种催化机制的可塑性在遗传密码进化的残余和现代重新编程遗传密码的努力（例如，核糖体掺入非规范氨基酸、核糖体酯形成）中都被观察到。然而，核糖体介导的聚合的局限性尚未得到充分探索。在这里，我们证明了通过活化的γ-酮和α-肼酯单体之间的环缩合反应，核糖体介导的哒嗪酮键聚合，而不是肽键。此外，我们还证明了由多个序列定义的交替哒嗪酮键组成的肽杂合寡聚物的核糖体催化合成。我们的结果强调了核糖体古老的成键机制的可塑性，扩大了核糖体可以合成的非规范聚合物主链的范围，并为合成生物学的新应用打开了大门。核糖体已进化为将 L-α-氨基酸聚合成包含肽骨架的蛋白质。在这里，使用核糖体在体外形成哒嗪酮主链，产生各种序列定义的交替嵌段共聚物。
The ribosome is a macromolecular machine that catalyzes the sequence-defined polymerization of L-α-amino acids into polypeptides. The catalysis of peptide bond formation between amino acid substrates is based on entropy trapping, wherein the adjacency of transfer RNA (tRNA)-coupled acyl bonds in the P-site and the α-amino groups in the A-site aligns the substrates for coupling. The plasticity of this catalytic mechanism has been observed in both remnants of the evolution of the genetic code and modern efforts to reprogram the genetic code (e.g., ribosomal incorporation of non-canonical amino acids, ribosomal ester formation). However, the limits of ribosome-mediated polymerization are underexplored. Here, rather than peptide bonds, we demonstrate ribosome-mediated polymerization of pyridazinone bonds via a cyclocondensation reaction between activated γ-keto and α-hydrazino ester monomers. In addition, we demonstrate the ribosome-catalyzed synthesis of peptide-hybrid oligomers composed of multiple sequence-defined alternating pyridazinone linkages. Our results highlight the plasticity of the ribosome’s ancient bond-formation mechanism, expand the range of non-canonical polymeric backbones that can be synthesized by the ribosome, and open the door to new applications in synthetic biology. Ribosomes have evolved to polymerize L-α-amino acids into proteins comprising a peptide backbone. Here, a pyridazinone backbone is formed using ribosomes in vitro, producing a variety of sequence-defined alternating block-copolymers.
登录
查看更多内容
影响因子：
14.9
作者：
Katoh T;Iwane Y;Suga H
通讯作者：
Suga H
影响因子：
18.2
作者：
Ad, Omer;Hoffman, Kyle S.;Schepartz, Alanna
通讯作者：
Schepartz, Alanna
影响因子：
4.7
作者：
Hong, Seok Hoon;Ntai, Ioanna;Haimovich, Adrian D.;Kelleher, Neil L.;Isaacs, Farren J.;Jewett, Michael C.
通讯作者：
Jewett, Michael C.
影响因子：
14.8
作者：
Kim, Do Soon;Watkins, Andrew;Bidstrup, Erik;Lee, Joongoo;Topkar, Ved;Kofman, Camila;Schwarz, Kevin J.;Liu, Yan;Pintilie, Grigore;Roney, Emily;Das, Rhiju;Jewett, Michael C.
通讯作者：
Jewett, Michael C.
影响因子：
15
作者：
Katoh, Takayuki;Suga, Hiroaki
通讯作者：
Suga, Hiroaki