Theory for measuring bivalent surface binding kinetics using total internal reflection with fluorescence photobleaching recovery

Total internal reflection with fluorescence photobleaching recovery (TIR-FPR) is a method for experimentally examining coupled diffusion and reaction kinetics at surfaces. In a previous work (Thompson et al. 1981. Biophys. J. 33:435–454), a theoretical basis for interpreting TIR-FPR data was describ...

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Bibliographic Details
Published in:Biophysical journal 1994-03, Vol.66 (3), p.898-911
Main Authors: Hsieh, H.V., Thompson, N.L.
Format: Article
Language:English
Subjects:
Animals
Binding Sites
Biophysical Phenomena
Biophysics
Cell Membrane - metabolism
Diffusion
Fluorescence
Kinetics
Membrane Proteins - metabolism
Membranes, Artificial
Models, Biological
Photochemistry
Surface Properties
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Summary:Total internal reflection with fluorescence photobleaching recovery (TIR-FPR) is a method for experimentally examining coupled diffusion and reaction kinetics at surfaces. In a previous work (Thompson et al. 1981. Biophys. J. 33:435–454), a theoretical basis for interpreting TIR-FPR data was described for monovalent ligands that undergo a reversible reaction with monovalent surface sites in a single step. Here, the theory for TIR-FPR has been extended to two different surface binding mechanisms that involve sequential, bivalent surface attachment. Methods for obtaining the intrinsic surface association and dissociation kinetic rates from measured fluorescence photobleaching recovery curves are described. The new theory should be applicable to the association of bivalent protein ligands such as antibodies with supported planar model membranes.
ISSN:0006-3495
1542-0086
DOI:10.1016/S0006-3495(94)80866-5