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Measuring Receptor–Ligand Binding Kinetics on Cell Surfaces: From Adhesion Frequency to Thermal Fluctuation Methods

  • Molecular Interactions
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Abstract

Interactions between surface-anchored receptors and ligands mediate cell–cell and cell–environment communications in many biological processes. Molecular interactions across two apposing cell membrane are governed by two-dimensional (2D) kinetics, which are physically distinct from and biologically more relevant than three-dimensional (3D) kinetics with at least one interacting molecular species in the fluid phase. Here we review two assays for measuring 2D binding kinetics: the adhesion frequency assay and the thermal fluctuation assay. The former measures the binding frequency as a function of contact duration and extracts the force-free 2D kinetics parameters by nonlinearly fitting the data with a probabilistic model. The latter detects bond formation/dissociation by monitoring the reduction/resumption of thermal fluctuations of a force sensor. Both assays are mechanically based and operate at the level of mostly single molecular interaction, which requires ultrasensitive force techniques. Characterization of one such technique, the biomembrane force probe, is presented.

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Acknowledgments

We thank our coworkers of references Chen et al. 1 and Huang et al. 5 who contributed the original data that are discussed here. This work was supported by National Institutes of Health Grants AI38282, AI44902, and HL091020. W.C. is a Predoctoral Fellowship recipient of the American Heart Association (Greater Southeast Affiliate).

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Author notes

  1. Krishna K. Sarangapani

    Present address: Woodruff School of Mechanical Engineering, Georgia Institute of Technology, Atlanta, GA, 30332, USA

Authors and Affiliations

  1. Woodruff School of Mechanical Engineering, Georgia Institute of Technology, Atlanta, GA, 30332, USA

    Wei Chen & Cheng Zhu

  2. Coulter Department of Biomedical Engineering, Georgia Institute of Technology, Atlanta, GA, 30332, USA

    Veronika I. Zarnitsyna, Krishna K. Sarangapani, Jun Huang & Cheng Zhu

Authors
  1. Wei Chen
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  2. Veronika I. Zarnitsyna
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  3. Krishna K. Sarangapani
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  4. Jun Huang
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  5. Cheng Zhu
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Correspondence to Cheng Zhu.

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Chen, W., Zarnitsyna, V.I., Sarangapani, K.K. et al. Measuring Receptor–Ligand Binding Kinetics on Cell Surfaces: From Adhesion Frequency to Thermal Fluctuation Methods. Cel. Mol. Bioeng. 1, 276–288 (2008). https://doi.org/10.1007/s12195-008-0024-8

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  • DOI: https://doi.org/10.1007/s12195-008-0024-8

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