Norris Vic, Krylov Sergey N, Agarwal Pratul K, White Glenn J
Theoretical Biology Unit, EA 4312, Department of Biology, University of Rouen, Mont Saint Aignan, France.
J Mol Microbiol Biotechnol. 2017;27(2):117-127. doi: 10.1159/000464443. Epub 2017 Apr 28.
The construction of switchable, radiation-controlled, aptameric enzymes - "swenzymes" - is, in principle, feasible. We propose a strategy to make such catalysts from 2 (or more) aptamers each selected to bind specifically to one of the substrates in, for example, a 2-substrate reaction. Construction of a combinatorial library of candidate swenzymes entails selecting a set of a million aptamers that bind one substrate and a second set of a million aptamers that bind the second substrate; the aptamers in these sets are then linked pairwise by a linker, thus bringing together the substrates. In the presence of the substrates, some linked aptamer pairs catalyze the reaction when exposed to external energy in the form of a specific frequency of low-intensity, nonionizing electromagnetic or acoustic radiation. Such swenzymes are detected via a separate product-capturing aptamer that changes conformation on capturing the product; this altered conformation allows it (1) to bind to every potential swenzyme in its vicinity (thereby giving a higher probability of capture to the swenzymes that generate the product) and (2) to bind to a sequence on a magnetic bead (thereby permitting purification of the swenzyme plus product-capturing aptamer by precipitation). Attempts to implement the swenzyme strategy may help elucidate fundamental problems in enzyme catalysis.
可切换的、辐射控制的适体酶——“开关酶”——的构建原则上是可行的。我们提出了一种策略,用2种(或更多)适体来制备这种催化剂,每种适体都被选择用于特异性结合例如双底物反应中其中一种底物。构建候选开关酶的组合文库需要选择一组一百万个能结合一种底物的适体和另一组一百万个能结合第二种底物的适体;然后这些组中的适体通过连接子两两相连,从而使底物聚集在一起。在有底物存在的情况下,一些相连的适体对在暴露于特定频率的低强度非电离电磁或声辐射形式的外部能量时会催化反应。这种开关酶通过一种单独的产物捕获适体来检测,该适体在捕获产物时会改变构象;这种改变的构象使其能够(1)结合其附近的每一种潜在开关酶(从而增加捕获产生该产物的开关酶的概率)以及(2)结合磁珠上的一个序列(从而允许通过沉淀纯化开关酶加产物捕获适体)。实施开关酶策略的尝试可能有助于阐明酶催化中的基本问题。