Insights into the ribosome function from the structures of non-arrested ribosome-nascent chain complexes.
Insights into the ribosome function from the structures of non-arrested ribosome-nascent chain complexes.
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DOI：
10.1038/s41557-022-01073-1
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发表时间：
2023-01
期刊：
影响因子：
21.8
通讯作者：
中科院分区：
文献类型：
--
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在蛋白质合成过程中，生长的多肽穿过核糖体出口隧道，并通过自身或通过感知出现在隧道中的各种小分子，如代谢物或抗生素来调节核糖体的活动。虽然核糖体-新生链状复合体(RNCC)的结构已经得到了广泛的研究，但由于缺乏一种大规模制备多肽合成的中间体肽-tRNAs的简单方法，这意味着很少有人关注代表核糖体功能活性状态的RNCCs。在这里，我们报道了通过一种基于天然化学连接的化学酶方法简便地合成稳定连接的多肽-tRNAs，并用它们来确定处于多肽转移酶中心(PTC)攻击前功能状态的RNCCs的几个结构。这些结构揭示了生长肽的C末端部分采用相同的统一β链构象，该构象由错综复杂的H-键网络与普遍保守的23SRRNA核苷酸稳定，并使核糖体如何以可比较的效率合成具有不同序列的生长肽。合成肽-tRNAs是具有挑战性的，因为没有酶可以直接将所需的多肽连接到tRNA上。目前已有研究表明，基于天然化学连接的化学酶方法可用于半合成肽-tRNAs，用于研究滞留和非滞留核糖体复合体的结构和生化。
During protein synthesis, the growing polypeptide threads through the ribosomal exit tunnel and modulates ribosomal activity by itself or by sensing various small molecules, such as metabolites or antibiotics, appearing in the tunnel. While arrested ribosome-nascent chain complexes (RNCCs) have been extensively studied structurally, the lack of a simple procedure for the large-scale preparation of peptidyl-tRNAs, intermediates in polypeptide synthesis that carry the growing chain, means that little attention has been given to RNCCs representing functionally active states of the ribosome. Here we report the facile synthesis of stably linked peptidyl-tRNAs—through a chemoenzymatic approach based on native chemical ligation—and use them to determine several structures of RNCCs in the functional pre-attack state of the peptidyl transferase center (PTC). These structures reveal that C-terminal parts of the growing peptides adopt the same uniform β-strand conformation stabilized by the intricate network of H-bonds with the universally conserved 23S rRNA nucleotides and rationalize how the ribosome synthesizes growing peptides having various sequences with comparable efficiencies. Synthesis of peptidyl-tRNAs is challenging because there are no enzymes that can directly attach the desired peptide to tRNA. Now it has been shown that a chemoenzymatic approach based on native chemical ligation can be used for the semi-synthesis of peptidyl-tRNAs for structural/biochemical studies of arrested and non-arrested ribosome complexes.
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影响因子：
16.8
作者：
Cabrita LD;Cassaignau AME;Launay HMM;Waudby CA;Wlodarski T;Camilloni C;Karyadi ME;Robertson AL;Wang X;Wentink AS;Goodsell L;Woolhead CA;Vendruscolo M;Dobson CM;Christodoulou J
通讯作者：
Christodoulou J
影响因子：
28.3
作者：
Herrero del Valle, Alba;Seip, Britta;Innis, C. Axel
通讯作者：
Innis, C. Axel
影响因子：
56.9
作者：
DAWSON, PE;MUIR, TW;KENT, SBH
通讯作者：
KENT, SBH
影响因子：
15
作者：
Doerfel, Lili K.;Wohlgemuth, Ingo;Rodnina, Marina V.
通讯作者：
Rodnina, Marina V.
影响因子：
11.4
作者：
Heurgué-Hamard, V;Karimi, R;Buckingham, RH
通讯作者：
Buckingham, RH