Development of a novel, high-affinity ssDNA trypsin inhibitor. (1st January 2019)

• Record Type: Journal Article
• Authors: Malicki, Stanislaw; Ksiazek, Miroslaw; Majewski, Pawel; Pecak, Aleksandra; Mydel, Piotr; Grudnik, Przemyslaw; Dubin, Grzegorz
• Is Part Of: Journal of enzyme inhibition and medicinal chemistry. Volume 34:Number 1(2019)
• Page Start: 638
• Page End: 643
• Publication Date: 2019-01-01
• Subjects: Aptamer -- ssDNA -- trypsin -- protease inhibitor; Enzyme inhibitors -- Periodicals; Enzyme Inhibitors -- periodicals; Biochemistry -- periodicals; 572.7
• DOI: 10.1080/14756366.2019.1569648
• Languages: English
• ISSNs: 1475-6366

Abstract

Abstract: 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 Ki 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.