title: Single molecule binding dynamics measured with atomic force microscopy.
abstract: We present a new method to analyse simultaneous Topography and RECognition Atomic Force Microscopy data such that it becomes possible to measure single molecule binding rates of surface bound proteins. We have validated this method on a model system comprising a S-layer surface modified with Strep-tagII for binding sites and strep-tactin bound to an Atomic Force Microscope tip through a flexible Poly-Ethylene-Glycol linker. At larger distances, the binding rate is limited by the linker, which limits the diffusion of the strep-tactin molecule, but at lateral distances below 3 nm, the binding rate is solely determined by the intrinsic molecular characteristics and the surface geometry and chemistry of the system. In this regime, Kon as determined from single molecule TREC data is in agreement with Kon determined using traditional biochemical methods.
authors.affiliation: Kamerlingh Onnes Laboratory, Leiden University, 2300 RA, Leiden, The Netherlands. Electronic address: es@physics.leidenuniv.nl.
authors.forename: M H
authors.initials: MH
authors.lastname: van Es
authors.affiliation: Institute of Biophysics, JKU Life Science Center O, Gruberstrae 40, A-4020 Linz, Austria. Electronic address: jltang@ciac.jl.cn.
authors.forename: J
authors.initials: J
authors.lastname: Tang
authors.affiliation: Center for Advanced Bioanalysis GmbH, Gruberstrasse 40, 4020 Linz, Austria. Electronic address: johannes.preiner@jku.at.
authors.forename: J
authors.initials: J
authors.lastname: Preiner
authors.affiliation: Institute of Biophysics, JKU Life Science Center O, Gruberstrae 40, A-4020 Linz, Austria. Electronic address: peter.hinterdorfer@jku.at.
authors.forename: P
authors.initials: P
authors.lastname: Hinterdorfer
authors.affiliation: Kamerlingh Onnes Laboratory, Leiden University, 2300 RA, Leiden, The Netherlands. Electronic address: oosterkamp@physics.leidenuniv.nl.
authors.forename: T H
authors.initials: TH
authors.lastname: Oosterkamp
mesh_terms: D001407:Bacillus / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism; D001665:Binding Sites; D007091:Image Processing, Computer-Assisted / Q000379:methods / Q000706:statistics & numerical data; D007700:Kinetics; D018625:Microscopy, Atomic Force / Q000379:methods* / Q000706:statistics & numerical data; D009004:Monosaccharide Transport Proteins / Q000737:chemistry / Q000378:metabolism; D009842:Oligopeptides / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D013499:Surface Properties
chemical_list: D001426:Bacterial Proteins; D009004:Monosaccharide Transport Proteins; D009842:Oligopeptides; C120455:SbpA protein, bacteria; C583458:Trp-Ser- His-Pro-Gln-Phe-Glu-Lys
keywords: AFM; Binding kinetics; Interaction; Simultaneous Topography and RECognition Imaging; Single molecule