Meli Marc, Vergne Jacques, Décout Jean-Luc, Maurel Marie-Christine
Institut Jacques Monod, Laboratoire de Biochimie de l'Evolution et Adaptabilité Moléculaire, Tour 43, 2 place Jussieu, 75251 Paris Cedex 05, France.
J Biol Chem. 2002 Jan 18;277(3):2104-11. doi: 10.1074/jbc.M107130200. Epub 2001 Nov 8.
RNA aptamers that are able to complex free adenine have been isolated by a SELEX (systematic evolution of ligands by exponential enrichment) procedure. The adenine binding site was revealed by sequence alignment for a prevalent cluster of aptamers, and its structure and interactions with adenine were probed by RNase digestion studies, lead cleavage, boundary determination experiments, and truncated sequences studies. A new purine binding motif was functionally and structurally characterized and compared with other RNAs specific to purine or adenylated compounds. The affinity for adenine and the specificity for other related targets were quantified. This work suggests that the adenine binding site is composed of two independent secondary structure elements forming a bipartite binding site that interacts with adenine in a new mode of purine recognition. Such binding is of great interest because the imidazole moiety is not trapped in the binding site, and would easily be available for catalytic activity.
能够与游离腺嘌呤形成复合物的RNA适体已通过SELEX(指数富集配体系统进化)程序分离出来。通过对一组普遍存在的适体进行序列比对揭示了腺嘌呤结合位点,并通过核糖核酸酶消化研究、铅离子切割、边界确定实验和截短序列研究对其结构以及与腺嘌呤的相互作用进行了探究。一种新的嘌呤结合基序在功能和结构上得到了表征,并与其他针对嘌呤或腺苷化化合物的RNA进行了比较。对腺嘌呤的亲和力以及对其他相关靶标的特异性进行了量化。这项工作表明,腺嘌呤结合位点由两个独立的二级结构元件组成,形成了一个二分结合位点,该位点以一种新的嘌呤识别模式与腺嘌呤相互作用。这种结合非常有趣,因为咪唑部分没有被困在结合位点中,并且易于用于催化活性。