title: Studies of the binding mechanism between aptamers and thrombin by circular dichroism, surface plasmon resonance and isothermal titration calorimetry.
abstract: Thrombin, a multifunctional serine protease, has both procoagulant and anticoagulant functions in human blood. Thrombin has two electropositive exosites. One is the fibrinogen-binding site and the other is the heparin-binding site. Over the past decade, two thrombin-binding aptamers (15-mer and 29-mer) were reported by SELEX technique. Recently, many studies examined the interactions between the 15-mer aptamer and thrombin extensively, but the data on the difference of these two aptamers binding to thrombin are still lacking and worth investigating for fundamental understanding. In the present study, we combined conformational data from circular dichroism (CD), kinetics and thermodynamics information from surface plasmon resonance (SPR) and isothermal titration calorimetry (ITC) to compare the binding mechanism between the two aptamers with thrombin. Special attentions were paid to the formation of G-quadruplex and the effects of ions on the aptamer conformation on the binding and the kinetics discrimination between specific and nonspecific interactions of the binding. The results indicated reasonably that the 15-mer aptamer bound to fibrinogen-binding site of thrombin using a G-quadruplex structure and was dominated by electrostatic interactions, while the 29-mer aptamer bound to heparin-binding site thrombin using a duplex structure and was driven mainly by hydrophobic effects.
authors: Institute of Systems Biology and Bioinformatics, National Central University, Jhong-Li, 320 Taiwan.
authors: Po-Hsun
authors: PH
authors: Lin
authors: Ren-Hao
authors: RH
authors: Chen
authors: Chung-Han
authors: CH
authors: Lee
authors: Yung
authors: Y
authors: Chang
authors: Chien-Sheng
authors: CS
authors: Chen
authors: Wen-Yih
authors: WY
authors: Chen
mesh_terms: D052157:Aptamers, Nucleotide / Q000737:chemistry* / Q000378:metabolism*; D002151:Calorimetry / Q000379:methods*; D002942:Circular Dichroism / Q000379:methods*; D005340:Fibrinogen / Q000378:metabolism; D054856:G-Quadruplexes; D006493:Heparin / Q000378:metabolism; D006801:Humans; D008958:Models, Molecular; D008968:Molecular Conformation; D020349:Surface Plasmon Resonance / Q000379:methods*; D013917:Thrombin / Q000737:chemistry* / Q000378:metabolism*
chemical_list: D052157:Aptamers, Nucleotide; C081676:thrombin aptamer; D005340:Fibrinogen; D006493:Heparin; D013917:Thrombin