Hohmura Ken I, Shi Hua, Hirayoshi Kazunori
Department of Molecular and Cellular Biology, Institute for Frontier Medical Sciences, Kyoto University.
Biosci Biotechnol Biochem. 2013;77(8):1739-46. doi: 10.1271/bbb.130296. Epub 2013 Aug 7.
Control of interactions among proteins is critical in the treatment of diseases, but the specificity required is not easily incorporated into small molecules. Macromolecules could be more suitable as antagonists in this situation, and RNA aptamers have become particularly promising. Here we describe a novel selection procedure for RNA aptamers against a protein that constitutes a single structural domain, the Drosophila TATA-binding protein (TBP). In addition to the conventional filter partitioning method with free TBP as target, we performed another experiment, in which the TATA-bound form of TBP was targeted. Aptamers generated by both selections were able to bind specifically to TBP, but the two groups showed characteristics which were clearly different in terms of their capability to compete with TATA-DNA, their effects on the TATA-bound form of TBP, and their effects on in vitro transcription. The method used to generate these two groups of aptamers can be used with other targets to direct aptamer specificity to discrete sites on the surface of a protein.
蛋白质间相互作用的调控在疾病治疗中至关重要，但所需的特异性难以融入小分子中。在这种情况下，大分子可能更适合作为拮抗剂，而RNA适配体已变得特别有前景。在此，我们描述了一种针对构成单一结构域的蛋白质——果蝇TATA结合蛋白（TBP）——筛选RNA适配体的新方法。除了以游离TBP为靶点的传统滤膜分区法外，我们还进行了另一项实验，其中以TBP与TATA结合的形式为靶点。通过这两种筛选方法产生的适配体都能够特异性结合TBP，但两组在与TATA-DNA竞争的能力、对TBP与TATA结合形式的影响以及对体外转录的影响方面表现出明显不同的特性。用于产生这两组适配体的方法可用于其他靶点，以将适配体特异性导向蛋白质表面的离散位点。
