Studies of the binding mechanism between aptamers and thrombin by circular dichroism, surface plasmon resonance and isothermal titration calorimetry

NCUIR > Engineering College > Department of Chemical and Materials Engineering > journal & Dissertation > Item 987654321/49973

Please use this identifier to cite or link to this item: http://ir.lib.ncu.edu.tw/handle/987654321/49973

Title:	Studies of the binding mechanism between aptamers and thrombin by circular dichroism, surface plasmon resonance and isothermal titration calorimetry
Authors:	Lin,PH;Chen,RH;Lee,CH;Chang,Y;Chen,CS;Chen,WY
Contributors:	化學工程與材料工程學系
Keywords:	SINGLE-STRANDED-DNA;SELECTION;PROTEINS;AFFINITY;SYSTEM;ALPHA;ASSAY
Date:	2011
Issue Date:	2012-03-27 16:27:47 (UTC+8)
Publisher:	國立中央大學
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-met aptamer bound to heparin-binding site thrombin using a duplex structure and was driven mainly by hydrophobic effects. (C) 2011 Elsevier B.V. All rights reserved.
Relation:	COLLOIDS AND SURFACES B-BIOINTERFACES
Appears in Collections:	[Department of Chemical and Materials Engineering] journal & Dissertation

Files in This Item:

File	Description	Size	Format
index.html		0Kb	HTML	648	View/Open

社群 sharing

Loading...