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In JoVE (1)
Other Publications (1)
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Articles by Tom Seegar in JoVE
Imaging eiwit-eiwit interacties In vivo
Tom Seegar, William Barton
Biochemistry and Molecular Biology, Virginia Commonwealth University
Dit protocol wordt beschreven hoe u foto eiwit-eiwit interacties met behulp van een FRET-gebaseerde nabijheid assay.
Other articles by Tom Seegar on PubMed
Molecular Cell. Mar, 2010 | Pubmed ID: 20227369
The Tie family of endothelial-specific receptor tyrosine kinases is essential for cell proliferation, migration, and survival during angiogenesis. Despite considerable similarity, experiments with Tie1- or Tie2-deficient mice highlight distinct functions for these receptors in vivo. The Tie2 receptor is further unique with respect to its structurally homologous ligands. Angiopoietin-2 and -3 can function as agonists or antagonists; angiopoietin-1 and -4 are constitutive agonists. To address the role of Tie1 in angiopoietin-mediated Tie2 signaling and determine the basis for the behavior of the individual angiopoietins, we used an in vivo FRET-based proximity assay to monitor Tie1 and -2 localization and association. We provide evidence for Tie1-Tie2 complex formation on the cell surface and identify molecular surface areas essential for receptor-receptor recognition. We further demonstrate that the Tie1-Tie2 interactions are dynamic, inhibitory, and differentially modulated by angiopoietin-1 and -2. Based on the available data, we propose a unified model for angiopoietin-induced Tie2 signaling.