A GENERAL-METHOD FOR SITE-SPECIFIC INCORPORATION OF UNNATURAL AMINO-ACIDS INTO PROTEINS
A GENERAL-METHOD FOR SITE-SPECIFIC INCORPORATION OF UNNATURAL AMINO-ACIDS INTO PROTEINS
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DOI：
10.1126/science.2649980
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发表时间：
1989-04-14
期刊：
影响因子：
56.9
通讯作者：
SCHULTZ, PG
中科院分区：
文献类型：
--
作者：
NOREN, CJ;ANTHONYCAHILL, SJ;SCHULTZ, PG
一种新的方法已经被开发出来，可以将非天然氨基酸特异地结合到蛋白质中。合成氨基酸被掺入酶中。-内酰胺酶通过使用化学上酰化的抑制转移RNA插入氨基酸，以响应取代编码感兴趣残基的密码子的停止密码子。肽定位将插入的氨基酸定位为单个肽，并且可以产生足够的酶进行纯化以达到均匀性。研究了几种转化型Phe66突变体的催化性能。有选择地用多种结构和电子变体替代蛋白质中的氨基酸的能力应该提供对蛋白质结构和功能的更详细的了解。
A new method has been developed that makes it possible to site-specifically incorporate unnatural amino acids into proteins. Synthetic amino acids were incorporated into the enzyme .beta.-lactamase by the use of a chemically acylated suppressor transfer RNA that inserted the amino acid in response to a stop codon substituted for the codon encoding residue of interest. Peptide mapping localized the inserted amino acid to a single peptide, and enough enzyme could be generated for purification to homogeneity. The catalytic properties of several mutants at the conversed Phe66 were characterized. The ability to selectively replace amino acids in a protein with a wide variety of structural and electronic variants should provide a more detailed understanding of protein structure and function.