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Real-Time Single Molecule Visualization of SH2 Domain Membrane Recruitment in Growth Factor Stimulated Cells

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SH2 Domains

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1555))

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Abstract

In the last decade, single molecule tracking (SMT) techniques have emerged as a versatile tool for molecular cell biology research. This approach allows researchers to monitor the real-time behavior of individual molecules in living cells with nanometer and millisecond resolution. As a result, it is possible to visualize biological processes as they occur at a molecular level in real time. Here we describe a method for the real-time visualization of SH2 domain membrane recruitment from the cytoplasm to epidermal growth factor (EGF) induced phosphotyrosine sites on the EGF receptor. Further, we describe methods that utilize SMT data to define SH2 domain membrane dynamics parameters such as binding (τ), dissociation (k d), and diffusion (D) rates. Together these methods may allow us to gain greater understanding of signal transduction dynamics and the molecular basis of disease-related aberrant pathways.

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Acknowledgements

I would like to thank Ji Yu, Joshua Jadwin, and Mari Ikeda for assistance with editing the manuscript.

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Authors and Affiliations

  1. Cell Analysis and Modeling, University of Connecticut Health Center, Farmington, CT, 06030, USA

    Dongmyung Oh

  2. Mechanobiology Institute, National University of Singapore, Singapore, Singapore

    Dongmyung Oh

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  1. Dongmyung Oh
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Correspondence to Dongmyung Oh .

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Editors and Affiliations

  1. Department of Genetics and Genome Sciences, University of Connecticut School of Medicine, Farmington, Connecticut, USA

    Kazuya Machida

  2. Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA

    Bernard A. Liu

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Oh, D. (2017). Real-Time Single Molecule Visualization of SH2 Domain Membrane Recruitment in Growth Factor Stimulated Cells. In: Machida, K., Liu, B. (eds) SH2 Domains. Methods in Molecular Biology, vol 1555. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6762-9_29

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  • DOI: https://doi.org/10.1007/978-1-4939-6762-9_29

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6760-5

  • Online ISBN: 978-1-4939-6762-9

  • eBook Packages: Springer Protocols

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