Laboratory of Proteolysis and Post-translational Modification of Proteins, Malopolska Centre of Biotechnology, Jagiellonian University, Krakow, Poland; Department of Microbiology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Krakow, Poland.
Laboratory of Proteolysis and Post-translational Modification of Proteins, Malopolska Centre of Biotechnology, Jagiellonian University, Krakow, Poland; Department of Microbiology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Krakow, Poland.
J Biol Chem. 2023 Aug;299(8):104889. doi: 10.1016/j.jbc.2023.104889. Epub 2023 Jun 5.
Human neutrophil elastase (HNE) plays a pivotal role in innate immunity, inflammation, and tissue remodeling. Aberrant proteolytic activity of HNE contributes to organ destruction in various chronic inflammatory diseases including emphysema, asthma, and cystic fibrosis. Therefore, elastase inhibitors could alleviate the progression of these disorders. Here, we used the systematic evolution of ligands by exponential enrichment to develop ssDNA aptamers that specifically target HNE. We determined the specificity of the designed inhibitors and their inhibitory efficacy against HNE using biochemical and in vitro methods, including an assay of neutrophil activity. Our aptamers inhibit the elastinolytic activity of HNE with nanomolar potency and are highly specific for HNE and do not target other tested human proteases. As such, this study provides lead compounds suitable for the evaluation of their tissue-protective potential in animal models.
人中性粒细胞弹性蛋白酶 (HNE) 在先天免疫、炎症和组织重塑中发挥着关键作用。HNE 的异常蛋白水解活性导致多种慢性炎症性疾病中的器官破坏,包括肺气肿、哮喘和囊性纤维化。因此,弹性蛋白酶抑制剂可以减轻这些疾病的进展。在这里,我们使用指数富集的配体系统进化来开发特异性针对 HNE 的 ssDNA 适体。我们使用生化和体外方法,包括中性粒细胞活性测定,确定了设计抑制剂的特异性及其对 HNE 的抑制效力。我们的适体以纳摩尔效力抑制 HNE 的弹性蛋白酶活性,并且对 HNE 具有高度特异性,而不针对其他测试的人类蛋白酶。因此,这项研究提供了适合在动物模型中评估其组织保护潜力的先导化合物。