Anlyses of enzymatic reaction about glycoconjugates using Atomic Force Microscopy in single molecule level

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Atomic force microscopy (AFM) can detect forces as intermolecular interaction forces by force-curve measurements. For example, when a ligand is immobilized on a chip, and a receptor is immobilized on a plate, we can measure single-molecular interaction forces between two molecules. The distribution of glycolipid microdomain in biomembrane is difficult to observe in nanometer scale. We measured a distribution of glycolipid Gb3Ceramide (Gb3Cer) in lipid bilayer by AFM force mapping measurement using Verotoxin modified tip.In this study, we have analyzed interaction forces between a verotoxin that is a substance responsible for o-157 and Gb3 sugar molecules on a cell surface. The rupture forces were obtained by changing loading rates and we can get an effective bond length which is an important parameter for an analysis of interaction mechanisms. We detected single-molecular interaction forces, and we discussed about the interaction mechanism in more detail by an analysis of the forces.