设计性核酶：将tRNA特异性编入柔性酶中。

Designer ribozymes: programming the tRNA specificity into flexizyme.

作者信息

Ramaswamy Krishna, Saito Hirohide, Murakami Hiroshi, Shiba Kiyotaka, Suga Hiroaki

机构信息

Departments of Chemistry and Biological Sciences, University at Buffalo, State University of New York, Buffalo, NY 14260-3000, USA.

出版信息

J Am Chem Soc. 2004 Sep 22;126(37):11454-5. doi: 10.1021/ja046843y.

DOI:10.1021/ja046843y

PMID:15366888

Abstract

Fx3 is an artificial ribozyme with the ability to aminoacylate various tRNAs with phenylalanine and its nonnatural derivatives. Herein we report a simple strategy to build tRNA specificity into the generic Fx3, by appending to its 3'-end a tRNA-specific sequence (TSS), which is complementary to the acceptor stem of the cognate tRNA. This new designer ribozyme, referred to as Fx10, is able to recognize its cognate tRNA via a 10-base-pair interaction that is formed after the invasion of the tRNA acceptor stem by the TSS. We have demonstrated that Fx10 can aminoacylate its cognate tRNA with a high degree of specificity and also discriminate against the noncognate tRNAs. Because the tRNA specificity can be easily programmed into Fx10, it is a custom-made catalyst to generate nonnatural aminoacyl-tRNAs.

摘要

Fx3是一种人工核酶，能够用苯丙氨酸及其非天然衍生物对各种tRNA进行氨酰化。在此，我们报告了一种简单的策略，通过在其3'末端附加一个与同源tRNA的受体茎互补的tRNA特异性序列（TSS），将tRNA特异性引入通用的Fx3中。这种新的设计核酶，称为Fx10，能够通过TSS侵入tRNA受体茎后形成的10个碱基对的相互作用识别其同源tRNA。我们已经证明，Fx10能够以高度特异性对其同源tRNA进行氨酰化，并且还能区分非同源tRNA。由于tRNA特异性可以很容易地编入Fx10中，它是一种定制催化剂，用于生成非天然氨酰-tRNA。