a Malopolska Centre of Biotechnology , Jagiellonian University , Krakow , Poland.
b Department of Microbiology, Faculty of Biochemistry , Biophysics and Biotechnology, Jagiellonian University , Krakow , Poland.
J Enzyme Inhib Med Chem. 2019 Dec;34(1):638-643. doi: 10.1080/14756366.2019.1569648.
Inhibitors of serine proteases are not only extremely useful in the basic research but are also applied extensively in clinical settings. Using Systematic Evolution of Ligands by Exponential Enrichment (SELEX) approach we developed a family of novel, single-stranded DNA aptamers capable of specific trypsin inhibition. Our most potent candidate (T24) and its short version (T59) were thoroughly characterised in terms of efficacy. T24 and T59 efficiently inhibited bovine trypsin with K of 176 nM and 475 nM, respectively. Interestingly, in contrast to the majority of known trypsin inhibitors, the selected aptamers have superior specificity and did not interact with porcine trypsin or any human proteases tested. These included plasmin and thrombin characterised by trypsin-like substrate specificity. Our results demonstrate that SELEX may be successfully employed in the development of potent and specific DNA based protease inhibitors.
丝氨酸蛋白酶抑制剂不仅在基础研究中非常有用，而且在临床环境中也得到了广泛应用。我们使用指数富集的配体系统进化（SELEX）方法开发了一系列新型的单链 DNA 适体，能够特异性抑制胰蛋白酶。我们最有效的候选物（T24）及其短版本（T59）在功效方面进行了彻底的表征。T24 和 T59 分别以 176 nM 和 475 nM 的 K 值有效地抑制牛胰蛋白酶。有趣的是，与大多数已知的胰蛋白酶抑制剂不同，所选适体具有更高的特异性，并且不与猪胰蛋白酶或任何测试的人类蛋白酶相互作用。这些蛋白酶包括具有胰蛋白酶样底物特异性的纤溶酶和凝血酶。我们的结果表明，SELEX 可成功用于开发强效和特异性的基于 DNA 的蛋白酶抑制剂。
