Abstract
The activity and/or localization of receptor tyrosine kinases and phosphatases are controlled by binding to cell-surface or secreted ligands. Identification of ligands for receptor tyrosine phosphatases (RPTPs) is essential for understanding their in vivo functions during development and disease. Here we describe a novel in vivo method to identify ligands and binding partners for RPTPs by staining live-dissected Drosophila embryos. Live dissected embryos are incubated with RPTP fusion proteins to detect ligand binding in embryos. This method can be streamlined to perform large-scale screens for ligands as well as to search for embryonic phenotypes.
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References
Lim WA, Pawson T (2010) Phosphotyrosine signaling: evolving a new cellular communication system. Cell 142(5):661–667. doi:10.1016/j.cell.2010.08.023
Fox AN, Zinn K (2005) The heparan sulfate proteoglycan syndecan is an in vivo ligand for the Drosophila LAR receptor tyrosine phosphatase. Curr Biol 15(19):1701–1711. doi:10.1016/j.cub.2005.08.035
Johnson KG, Tenney AP, Ghose A, Duckworth AM, Higashi ME, Parfitt K, Marcu O, Heslip TR, Marsh JL, Schwarz TL, Flanagan JG, Van Vactor D (2006) The HSPGs Syndecan and Dallylike bind the receptor phosphatase LAR and exert distinct effects on synaptic development. Neuron 49(4):517–531. doi:10.1016/j.neuron.2006.01.026
Lee HK, Cording A, Vielmetter J, Zinn K (2013) Interactions between a receptor tyrosine phosphatase and a cell surface ligand regulate axon guidance and glial-neuronal communication. Neuron 78(5):813–826. doi:10.1016/j.neuron.2013.04.001
Wright AP, Fox AN, Johnson KG, Zinn K (2010) Systematic screening of Drosophila deficiency mutations for embryonic phenotypes and orphan receptor ligands. PLoS One 5(8):e12288. doi:10.1371/journal.pone.0012288
Ozkan E, Carrillo RA, Eastman CL, Weiszmann R, Waghray D, Johnson KG, Zinn K, Celniker SE, Garcia KC (2013) An extracellular interactome of immunoglobulin and LRR proteins reveals receptor-ligand networks. Cell 154(1):228–239. doi:10.1016/j.cell.2013.06.006
Lee HK, Wright AP, Zinn K (2009) Live dissection of Drosophila embryos: streamlined methods for screening mutant collections by antibody staining. J Vis Exp (34). doi:10.3791/1647
Brent JR, Werner KM, McCabe BD (2009) Drosophila larval NMJ dissection. J Vis Exp (24). doi:10.3791/1107
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Bali, N., Lee, HK.(., Zinn, K. (2016). Live Staining of Drosophila Embryos with RPTP Fusion Proteins to Detect and Characterize Expression of Cell-Surface RPTP Ligands. In: Pulido, R. (eds) Protein Tyrosine Phosphatases. Methods in Molecular Biology, vol 1447. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3746-2_20
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DOI: https://doi.org/10.1007/978-1-4939-3746-2_20
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-3744-8
Online ISBN: 978-1-4939-3746-2
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