Expansion of the Genetic Code Through the Use of Modified Bacterial Ribosomes.
Expansion of the Genetic Code Through the Use of Modified Bacterial Ribosomes.
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DOI：
10.1016/j.jmb.2021.167211
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发表时间：
2022-04-30
影响因子：
5.6
通讯作者：
Hecht, Sidney M.
中科院分区：
文献类型：
--
作者：
Hecht, Sidney M.
关键词：
生物蛋白质合成是由核糖体介导的，利用~20个蛋白原氨基酸作为构建单元。通过使用错酰化的tRNAs，目前可以通过几种策略中的任何一种来获得，现在可以利用体外和体内蛋白质生物合成来精制含有更多种类的氨基酸构建块的蛋白质。然而，这种更广泛的氨基酸的加入仅限于核糖体所利用的那些物种。因此，随着时间的推移，几乎所有使用的底物都是L-α-氨基酸。近年来，各种结构和生化研究为23S核糖体RNA中参与肽键形成的区域提供了重要的见解。随后的实验，包括对形成肽键所需的23S rRNA关键区域的随机化，提供了含有在关键区域修饰了rrnB基因的质粒的大肠杆菌文库。基于使用具有改变的氨基酸的改良的嘌呤霉素衍生物的选择，然后鉴定对单独的嘌呤霉素衍生物唯一敏感的克隆。这些克隆通常识别错酰化的tRNAs，其中含有类似于修饰的嘌呤霉素中的改变的氨基酸，并将氨基酸类似物结合到蛋白质中。以这种方式，可以实现含有D-氨基酸、β-氨基酸、磷酸化氨基酸的蛋白质的合成，以及其中亲核氨基不在α-位置的长链和环状氨基酸。特别令人感兴趣的是具有不同用途的二肽和二肽模拟物。
Biological protein synthesis is mediated by the ribosome, and employs ~20 proteinogenic amino acids as building blocks. Through the use of misacylated tRNAs, presently accessible by any of several strategies, it is now possible to employ in vitro and in vivo protein biosynthesis to elaborate proteins containing a much larger variety of amino acid building blocks. However, the incorporation of this broader variety of amino acids is limited to those species utilized by the ribosome. As a consequence, virtually all of the substrates utilized over time have been L-α-amino acids. In recent years, a variety of structural and biochemical studies have provided important insights into those regions of the 23S ribosomal RNA that are involved in peptide bond formation. Subsequent experiments, involving the randomization of key regions of 23S rRNA required for peptide bond formation, have afforded libraries of E. coli harboring plasmids with the rrnB gene modified in the key regions. Selections based on the use of modified puromycin derivatives with altered amino acids then identified clones uniquely sensitive to individual puromycin derivatives. These clones often recognized misacylated tRNAs containing altered amino acids similar to those in the modified puromycins, and incorporated the amino acid analogues into proteins. In this fashion, it has been possible to realize the synthesis of proteins containing D-amino acids, β-amino acids, phosphorylated amino acids, as well as long chain and cyclic amino acids in which the nucleophilic amino group is not in the α-position. Of special interest have been dipeptides and dipeptidomimetics of diverse utility.
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影响因子：
14.9
作者：
Fan C;Xiong H;Reynolds NM;Söll D
通讯作者：
Söll D
影响因子：
3.5
作者：
Burakovsky, Dmitry E.;Sergiev, Petr V.;Dontsova, Olga A.
通讯作者：
Dontsova, Olga A.
DOI：
10.1073/pnas.48.6.1086
发表时间：
1962-01-01
影响因子：
11.1
作者：
CHAPEVILLE, F;EHRENSTEIN, GV;LIPMANN, F
通讯作者：
LIPMANN, F
影响因子：
14.9
作者：
GREGORY, ST;LIEBERMAN, KR;DAHLBERG, AE
通讯作者：
DAHLBERG, AE
影响因子：
2.9
作者：
Dedkova, Larisa M.;Fahmi, Nour Eddine;Hecht, Sidney M.
通讯作者：
Hecht, Sidney M.