Single molecule binding dynamics measured with atomic force microscopy
Sprache des Titels:
Englisch
Original Kurzfassung:
WepresentanewmethodtoanalysesimultaneousTopographyandRECognitionAtomicForceMicroscopy
datasuchthatitbecomespossibletomeasuresinglemoleculebindingratesofsurfaceboundproteins.We
havevalidatedthismethodonamodelsystemcomprisingaS-layersurfacemodified with Strep-tagII for
bindingsitesandstrep-tactinboundtoanAtomicForceMicroscopetipthrougha flexiblePoly-Ethylene-
Glycollinker.Atlargerdistances,thebindingrateislimitedbythelinker,whichlimitsthediffusionofthe
strep-tactinmolecule,butatlateraldistancesbelow3nm,thebindingrateissolelydeterminedbythe
intrinsicmolecularcharacteristicsandthesurfacegeometryandchemistryofthesystem.Inthisregime, Kon
asdeterminedfromsinglemoleculeTRECdataisinagreementwith Kon determinedusingtraditional
biochemicalmethods.