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Biophys. J. BioFAST: First Published September 21, 2007. doi:10.1529/biophysj.107.117895
© 2007 by the Biophysical Society.


A more recent version of this article appeared on January 15, 2008.
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CELL BIOPHYSICS

Monitoring Receptor-ligand Interactions Between Surfaces by Thermal Fluctuations

Wei Chen 1, Evan A. Evans 2, Rodger P. McEver 3 and Cheng Zhu 1*

1 Georgia Institute of Technology
2 Boston University
3 Oklahoma Medical Research Foundation

* To whom correspondence should be addressed. E-mail: cheng.zhu{at}me.gatech.edu.

Submitted on July 23, 2007
Revised on August 13, 2007
Accepted on 31 August 2007


   Abstract
We describe a new method for determining receptor-ligand association/dissociation events across the interface of two surfaces (2D binding) by monitoring abrupt decrease/resumption in thermal fluctuations of a biomembrane force probe. Our method has been validated by rigorous control experiments and kinetic experiments. We show that cellular on-rate of association can be measured by analysis of intervals from a dissociation event to the next association event (waiting times). Similarly, off-rate of molecular dissociation can be measured by analysis of intervals from an association event to the next dissociation event (bond lifetimes). Different types of molecular bonds could be distinguished by different levels of reduction in thermal fluctuations. This novel method provides a powerful tool to study cell adhesion and signaling mediated by single or multiple receptor-ligand species.

Key Words: biomembrane force probe, kinetics, receptor ligand interactions, thermal fluctuations, two dimensional binding




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Copyright © 2007 by the Biophysical Society.