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Single-Molecule Kinetic Analysis of Receptor Protein Tyrosine Kinases

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Cell Signaling Reactions

Abstract

Signaling pathways mediated by receptor tyrosine kinases (RTKs) are among the most important pathways regulating various functions and behaviors in mammalian cells. Although many studies performed over several decades have revealed the molecular mechanisms underlying the cellular events regulated by these pathways, the overall structures of the pathways remain unclear, especially their quantitative properties. A technology has emerged that can potentially address these issues. Recent developments in optical microscopy and molecular biology allow us to visualize the behaviors of single RTK molecules and their association partners with fluorescent probes in living cells. Using the quantitative nature of these single-molecule measurements, we studied the signaling of epidermal growth factor (EGF) and nerve growth factor (NGF), both of which stimulate RTK systems. Single-molecule analyses revealed molecular dynamics and kinetics that cannot be demonstrated with conventional biochemical methods. These include the kinetic transitions of these receptors induced by ligand binding, signal amplification by the dynamic interactions between active and inactive receptors, downstream signaling with a memory effect exerted by the receptor molecule, and shifts in the motional modes of ligand-receptor complexes. These novel insights obtained from single-molecule studies suggest that detailed models of RTK signaling, which involve signal processing depend on protein dynamics.

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Acknowledgments

We would like to thank Hiroaki Takagi, Tomomi Tani, Tatsuo Shibata, Masahiro Ueda, Toshio Yanagida, and members of our laboratory for their collaboration, continual encouragement, and helpful discussions.

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  1. Cellular Informatics Laboratory, Advance Science Institute, RIKEN Hirosawa 2-1, Wako, Saitama, 351-0198, Japan

    Michio Hiroshima & Yasushi Sako

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  1. Michio Hiroshima
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  2. Yasushi Sako
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Correspondence to Michio Hiroshima .

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  1. , Cellular Informatics Laboratory, RIKEN, Hirosawa 2-1, Wako, 351-0198, Japan

    Yasushi Sako

  2. Graduate School of Frontier Biosciences, Laboratories of Nanobiology, Osaka University, Yamadaoka 1-3, Suita, 565-871, Japan

    Masahiro Ueda

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Hiroshima, M., Sako, Y. (2011). Single-Molecule Kinetic Analysis of Receptor Protein Tyrosine Kinases. In: Sako, Y., Ueda, M. (eds) Cell Signaling Reactions. Springer, Dordrecht. https://doi.org/10.1007/978-90-481-9864-1_1

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