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Single Molecule Microscopy in Neurobiology pp 115–129Cite as

Approaching Protein-Protein Interactions in Membranes Using Single-Particle Tracking and Packing Coefficient Analysis

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Part of the book series: Neuromethods ((NM,volume 154))

Abstract

Molecules diffuse randomly in the plane of the plasma membrane due to membrane fluidity. However, interactions with other molecules may introduce noticeable changes in diffusion behavior such as a slowdown or immobilization by formation of complexes. As a consequence, the analysis of transitions between diffusive behaviors can provide effective rate constants of molecular interactions. Tracking the positions of labeled single molecules, i.e., using single-particle tracking (SPT), is particularly pertinent for this kind of studies. We provide here a step-by-step protocol for SPT experiments and analyses needed to estimate the effective rate constants of molecular interactions. Classical SPT data analyses using the mean square displacement provide the average diffusive behavior, thus precluding the analysis of transitions. We propose an alternative approach to overcome this problem, namely the packing coefficient (Pc) analysis. We illustrate the application of this method to the interactions of neurotransmitter receptors with their scaffolding proteins.

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Acknowledgments

This work was supported by the Agence Nationale de la Recherche “Synaptune” (Programme blanc, ANR-12-BSV4-0019-01), the ERC advanced research grant “PlasltInhib,” the program “Investissements d’Avenir” (ANR-10-LABX-54 MEMOLIFE and ANR-11-IDEX-0001-02 PSL Research University), and the Institut National de la Santé et de la Recherche Médicale (INSERM). Authors declare no conflict of interest.

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Authors and Affiliations

  1. Institut du Fer à Moulin (IFM), INSERM, UMR-S 1270, Sorbonne Université, Paris, France

    Marianne Renner

  2. Institute of Biology of the École Normale Supérieure (IBENS), CNRS, INSERM, PSL Research University, Paris, France

    Antoine Triller

Authors
  1. Marianne Renner
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  2. Antoine Triller
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Corresponding author

Correspondence to Marianne Renner .

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Editors and Affiliations

  1. Laboratory of Cellular and Molecular Neurobiology, Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka, Japan

    Nobuhiko Yamamoto

  2. Laboratory for Cell Polarity Regulation, RIKEN Center for Biosystems Dynamics Research, Suita, Osaka, Japan

    Yasushi Okada

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Renner, M., Triller, A. (2020). Approaching Protein-Protein Interactions in Membranes Using Single-Particle Tracking and Packing Coefficient Analysis. In: Yamamoto, N., Okada, Y. (eds) Single Molecule Microscopy in Neurobiology . Neuromethods, vol 154. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0532-5_6

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  • DOI: https://doi.org/10.1007/978-1-0716-0532-5_6

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0531-8

  • Online ISBN: 978-1-0716-0532-5

  • eBook Packages: Springer Protocols

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