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Analysis of the interaction of Plexin-B1 and Plexin-B2 with Rnd family proteins

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  • Thomas Wylie
  • Ritu Garg
  • Anne J Ridley
  • Maria R Conte

Abstract

The Rnd family of proteins, Rnd1, Rnd2 and Rnd3, are atypical Rho family GTPases, which bind to but do not hydrolyse GTP. They interact with plexins, which are receptors for semaphorins, and are hypothesised to regulate plexin signalling. We recently showed that each Rnd protein has a distinct profile of interaction with three plexins, Plexin-B1, Plexin-B2 and Plexin-B3, in mammalian cells, although it is unclear which region(s) of these plexins contribute to this specificity. Here we characterise the binary interactions of the Rnd proteins with the Rho-binding domain (RBD) of Plexin-B1 and Plexin-B2 using biophysical approaches. Isothermal titration calorimetry (ITC) experiments for each of the Rnd proteins with Plexin-B1-RBD and Plexin-B2-RBD showed similar association constants for all six interactions, although Rnd1 displayed a small preference for Plexin-B1-RBD and Rnd3 for Plexin-B2-RBD. Furthermore, mutagenic analysis of Rnd3 suggested similarities in its interaction with both Plexin-B1-RBD and Plexin-B2-RBD. These results suggest that Rnd proteins do not have a clear-cut specificity for different Plexin-B-RBDs, possibly implying the contribution of additional regions of Plexin-B proteins in conferring functional substrate selection.

Suggested Citation

  • Thomas Wylie & Ritu Garg & Anne J Ridley & Maria R Conte, 2017. "Analysis of the interaction of Plexin-B1 and Plexin-B2 with Rnd family proteins," PLOS ONE, Public Library of Science, vol. 12(10), pages 1-13, October.
    • Handle: RePEc:plo:pone00:0185899
    DOI: 10.1371/journal.pone.0185899
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    References listed on IDEAS

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    1. Roberta Azzarelli & Emilie Pacary & Ritu Garg & Patricia Garcez & Debbie van den Berg & Philippe Riou & Anne J. Ridley & Roland H. Friedel & Maddy Parsons & François Guillemot, 2014. "An antagonistic interaction between PlexinB2 and Rnd3 controls RhoA activity and cortical neuron migration," Nature Communications, Nature, vol. 5(1), pages 1-12, May.
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    Cited by:

    1. Xiujie Ge & Lan Chen & Dexing Li & Renxiao Liu & Guanglu Ge, 2020. "Estimation of non-constant variance in isothermal titration calorimetry using an ITC measurement model," PLOS ONE, Public Library of Science, vol. 15(12), pages 1-15, December.

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