Institute of Advanced Energy, Kyoto University, Gokasho, Uji, Japan.
FEBS J. 2012 Apr;279(8):1456-63. doi: 10.1111/j.1742-4658.2012.08538.x. Epub 2012 Mar 16.
The structure of r(GGAGGAGGAGGA) (R12) changes from a single-stranded form to a compact quadruplex one in response to K(+). In a hammerhead ribozyme, two portions of the catalytic core are linked with the stem and are located in close proximity in order to exert activity. In this study, the stem was replaced by R12 (or R11, which lacks the terminal A residue) with or without linker residues. One of the newly constructed ribozymes exhibited enhanced activity in response to K(+), and we suggest that quadruplex formation restored the active catalytic core. Other ribozymes exhibited repressed activity in response to K(+), suggesting that formation of the active core was prevented. Thus, we have succeeded in developing 'intelligent' ribozymes whose activity is either repressed or enhanced in response to K(+). This switching capability may have therapeutic applications because of the differences between intra- and extracellular K(+) concentrations.
当钾离子存在时,r(GGAGGAGGAGGA)(R12)的结构从单链形式转变为紧凑的四链体形式。在锤头核酶中,催化核心的两个部分与茎相连,并紧密相邻以发挥活性。在这项研究中,用 R12(或缺少末端 A 残基的 R11)代替茎,带有或不带有连接残基。新构建的核酶之一对钾离子的反应表现出增强的活性,我们认为四链体的形成恢复了活性催化核心。其他核酶对钾离子的反应表现出抑制活性,表明活性核心的形成受到了阻碍。因此,我们成功地开发了“智能”核酶,其活性可以根据钾离子的存在与否而被抑制或增强。这种切换能力可能具有治疗应用价值,因为细胞内和细胞外钾离子浓度存在差异。