Noren C J, Anthony-Cahill S J, Griffith M C, Schultz P G
Department of Chemistry, University of California, Berkeley 94720.
Science. 1989 Apr 14;244(4901):182-8. doi: 10.1126/science.2649980.
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 conserved 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.
已开发出一种新方法，该方法能够将非天然氨基酸位点特异性地掺入蛋白质中。通过使用化学酰化的抑制性转移RNA，将合成氨基酸掺入β-内酰胺酶中，该转移RNA会响应取代编码目标残基密码子的终止密码子而插入氨基酸。肽图分析将插入的氨基酸定位到单个肽段，并且可以产生足够的酶以纯化至均一性。对保守的苯丙氨酸66处的几个突变体的催化特性进行了表征。用多种结构和电子变体选择性取代蛋白质中氨基酸的能力应能更详细地了解蛋白质的结构和功能。
