Hydrodynamic effects in fast AFM single-molecule force measurements.
Atomic force microscopy (AFM) allows the critical forces that unfold single proteins and rupture individual receptor-ligand bonds to be measured. To derive the shape of the energy landscape, the dynamic strength of the system is probed at different force loading rates. This is usually achieved by varying the pulling speed between a few nm/s and a few microm/s, although for a more complete investigation of the kinetic properties higher speeds are desirable. Above 10 microm/s, the hydrodynamic drag force acting on the AFM cantilever reaches the same order of magnitude as the molecular forces. This has limited the maximum pulling speed in AFM single-molecule force spectroscopy experiments. Here, we present an approach for considering these hydrodynamic effects, thereby allowing a correct evaluation of AFM force measurements recorded over an extended range of pulling speeds (and thus loading rates). To support and illustrate our theoretical considerations, we experimentally evaluated the mechanical unfolding of a multi-domain protein recorded at 30 microm/s pulling speed.
Janovjak
Harald
H
BioTechnological Center, University of Technology Dresden, 01307 Dresden, Germany.
Struckmeier
Jens
J
Müller
Daniel J
DJ
D000465:Algorithms; D003198:Computer Simulation; D004548:Elasticity; D044085:Microfluidics / Q000379:methods*; D018625:Microscopy, Atomic Force / Q000379:methods*; D008956:Models, Chemical; D008958:Models, Molecular; D010812:Physical Stimulation / Q000379:methods*; D011485:Protein Binding; D011506:Proteins / Q000032:analysis* / Q000737:chemistry*; D013314:Stress, Mechanical; D014783:Viscosity
D011506:Proteins