Monitoring ligand-receptor interactions by photonic force microscopy.
We introduce a method for the acquisition of single molecule force measurements of ligand-receptor interactions using the photonic force microscope (PFM). Biotin-functionalized beads, manipulated with an optical trap, and a streptavidin-functionalized coverslip were used to measure the effect of different pulling forces on the lifetime of individual streptavidin-biotin complexes. By optimizing the design of the optical trap and selection of the appropriate bead size, pulling forces in excess of 50 pN were achieved. Based on the amplitude of three-dimensional (3D) thermal position fluctuations of the attached bead, we were able to select for a bead-coverslip interaction that was mediated by a single streptavidin-biotin complex. Moreover, the developed experimental system was greatly accelerated by automation of data acquisition and analysis. In force-dependent kinetic measurements carried out between streptavidin and biotin, we observed that the streptavidin-biotin complex exhibited properties of a catch bond, with the lifetime increasing tenfold when the pulling force increased from 10 to 20 pN. We also show that silica beads were more appropriate than polystyrene beads for the force measurements, as tethers, longer than 200 nm, could be extracted from polystyrene beads.
Jeney
Sylvia
S
M E Müller Institute for Structural Biology, Biozentrum, University of Basel, Basel, Switzerland. sylvia.jeney@unibas.ch
Mor
Flavio
F
Koszali
Roland
R
Forró
László
L
Moy
Vincent T
VT
D001669:Biochemical Phenomena / Q000502:physiology*; D001696:Biomechanical Phenomena; D001710:Biotin / Q000737:chemistry / Q000378:metabolism; D002138:Calibration; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008024:Ligands; D018625:Microscopy, Atomic Force / Q000379:methods*; D008863:Microspheres; D017785:Photons; D011137:Polystyrenes; D011485:Protein Binding; D019809:Streptavidin / Q000737:chemistry / Q000378:metabolism; D013696:Temperature
D008024:Ligands; D011137:Polystyrenes; D001710:Biotin; D019809:Streptavidin