Sassanfar M, Szostak J W
Department of Genetics, Harvard Medical School, Boston, Massachusetts.
Nature. 1993 Aug 5;364(6437):550-3. doi: 10.1038/364550a0.
RNAs that contain specific high-affinity binding sites for small molecule ligands immobilized on a solid support are present at a frequency of roughly one in 10(10)-10(11) in pools of random sequence RNA molecules. Here we describe a new in vitro selection procedure designed to ensure the isolation of RNAs that bind the ligand of interest in solution as well as on a solid support. We have used this method to isolate a remarkably small RNA motif that binds ATP, a substrate in numerous biological reactions and the universal biological high-energy intermediate. The selected ATP-binding RNAs contain a consensus sequence, embedded in a common secondary structure. The binding properties of ATP analogues and modified RNAs show that the binding interaction is characterized by a large number of close contacts between the ATP and RNA, and by a change in the conformation of the RNA.
对于固定在固体支持物上的小分子配体而言,含有特定高亲和力结合位点的RNA在随机序列RNA分子库中的出现频率约为10的10次方至10的11次方分之一。在此,我们描述了一种新的体外筛选程序,旨在确保分离出既能在溶液中又能在固体支持物上结合目标配体的RNA。我们已使用此方法分离出一个非常小的RNA基序,它能结合ATP,ATP是众多生物反应中的底物以及通用的生物高能中间体。所筛选出的ATP结合RNA包含一个共有序列,该序列嵌入一个共同的二级结构中。ATP类似物和修饰RNA的结合特性表明,这种结合相互作用的特征是ATP与RNA之间存在大量紧密接触,并且RNA的构象发生了变化。