Functional and Structural Characterization of Nucleic Acid Ligands That Bind to Activated Coagulation Factor XIII

Abstract

Coagulation factor XIII (FXIII) is a protransglutaminase which plays an important role in clot stabilization and composition by cross-linking the α- and γ-chains of fibrin and increasing the resistance of the clot to mechanical and proteolytic challenges. In this study, we selected six DNA aptamers specific for activated FXIII (FXIIIa) and investigated the functional characterization of FXIIIa after aptamer binding. One of these aptamers, named FA12, efficiently captures FXIIIa even in the presence of zymogenic FXIII subunits. Furthermore, this aptamer inhibits the incorporation of FXIII and α2-antiplasmin (α2AP) into fibrin(ogen) with IC50-values of 38 nM and 17 nM, respectively. In addition to FA12, also another aptamer, FA2, demonstrated significant effects in plasma-based thromboelastometry (rotational thromboelastometry analysis, ROTEM)-analysis where spiking of the aptamers into plasma decreased clot stiffness and elasticity (p < 0.0001). The structure–function correlations determined by combining modeling/docking strategies with quantitative in vitro assays revealed spatial overlap of the FA12 binding site with the binding sites of two FXIII substrates, fibrinogen and α2AP, while FA2 binding sites only overlap those of fibrinogen. Taken together, these features especially render the aptamer FA12 as an interesting candidate molecule for the development of FXIIIa-targeting therapeutic strategies and diagnostic assays.

Thrombomodulin activation driven by LXR agonist attenuates renal injury in diabetic nephropathy

“…Inflammation and thrombosis are increasingly recognized as interrelated biological processes but their role in the pathogenesis of DN is still unclear ( 8 , 9 ). Due to the important role of TM and FXIII-A in inflammation and coagulation process, we aimed to reveal the interaction between FXIII-A and TM activation and their effects to against DN.…”

“…This accompanying inflammation leads to hypercoagulation and hypofibrinolysis, resulting in coagulopathies ( 4 ). Inflammation and thrombosis are increasingly recognized as interrelated biologic processes but their role in the pathogenesis of DN remains unclear ( 8 , 9 ).…”

Cardiac Troponin Biosensor Designs: Current Developments and Remaining Challenges

“…Moreover, the impressive results obtained for the detection of cardiac troponins support the further development and optimization of these biosensors for the detection of novel discovered diagnostic AMI biomarkers. The versatility of aptamers to selectively bind specific molecules could be exploited to realize diagnostic tools that are able to detect coagulation factors such as FXIII, which play a crucial role in the diagnosis and treatment of cardiac diseases [ 91 , 92 ]. The continuous effort invested in the design of diagnostic tools for AMI and cardiac diseases in general is impressive.…”

Aptamer Loaded Superparamagnetic Beads For Selective Capturing And Gentle Release of Activated Protein C

“…Aptamers are single-stranded synthetic oligonucleotides generated by an in vitro selection process termed systematic evolution of ligands by exponential enrichment (SELEX) 15,16 . Aptamers bind to their target molecules with high a nity and speci city and involve the advantage of discrimination between active and inactive structural conformation of an enzyme 17,18 . In the context of a nity separation, aptamers offer advantages over antibodies which suit them for designing aptamer-based a nity puri cation strategies.…”