Constitutive Association of Tie1 and Tie2 with Endothelial Integrins is Functionally Modulated by Angiopoietin-1 and Fibronectin.
Functional cross-talk between Tie2 and Integrin signaling pathways is essential to coordinate endothelial cell adhesion and migration in response to the extracellular matrix, yet the mechanisms behind this phenomenon are unclear. Here, we examine the possibility that receptor cross-talk is driven through uncharacterized Tie-integrin interactions on the endothelial surface. Using a live cell FRET-based proximity assay, we monitor Tie-integrin receptor recognition and demonstrate that both Tie1 and Tie2 readily associate with integrins α5ß1 and αVß3 through their respective ectodomains. Although not required, Tie2-integrin association is significantly enhanced in the presence of the extracellular component and integrin ligand fibronectin. In vitro binding assays with purified components reveal that Tie-integrin recognition is direct, and further demonstrate that the receptor binding domain of the Tie2 ligand Ang-1, but not the receptor binding domain of Ang-2, can independently associate with α5ß1 or αVß3. Finally, we reveal that cooperative Tie/integrin interactions selectively stimulate ERK/MAPK signaling in the presence of both Ang-1 and fibronectin, suggesting a molecular mechanism to sensitize Tie2 to extracellular matrix. We provide a mechanistic model highlighting the role of receptor localization and association in regulating distinct signaling cascades and in turn, the angiogenic switch.
Virginia Commonwealth University, Department of Biochemistry and Molecular Biology, Richmond, Virginia, 23298, United States of America.
Annamarie C
AC
Dalton
Ben-Gurion University of the Negev, Department of Biotechnology Engineering and the National Institute of Biotechnology in the Negev, Beer-Sheva, 8410501, Israel.
Tomer
T
Shlamkovitch
Ben-Gurion University of the Negev, Department of Biotechnology Engineering and the National Institute of Biotechnology in the Negev, Beer-Sheva, 8410501, Israel.
Niv
N
Papo
Virginia Commonwealth University, Department of Biochemistry and Molecular Biology, Richmond, Virginia, 23298, United States of America.
William A
WA
Barton
D042683:Angiopoietin-1 / Q000235:genetics* / Q000378:metabolism; D002448:Cell Adhesion / Q000235:genetics; D042783:Endothelial Cells / Q000378:metabolism; D005109:Extracellular Matrix / Q000235:genetics / Q000378:metabolism; D005353:Fibronectins / Q000235:genetics* / Q000378:metabolism; D031541:Fluorescence Resonance Energy Transfer; D006801:Humans; D039081:Integrin alpha5beta1 / Q000235:genetics* / Q000378:metabolism; D039302:Integrin alphaVbeta3 / Q000235:genetics* / Q000378:metabolism; D020935:MAP Kinase Signaling System / Q000235:genetics; D010766:Phosphorylation; D011485:Protein Binding; D042810:Receptor, TIE-1 / Q000235:genetics* / Q000378:metabolism; D042787:Receptor, TIE-2 / Q000235:genetics* / Q000378:metabolism; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics / Q000378:metabolism; D033921:Vesicular Transport Proteins / Q000235:genetics
C467630:ANGPT1 protein, human; D042683:Angiopoietin-1; C067625:FN1 protein, human; D005353:Fibronectins; D039081:Integrin alpha5beta1; D039302:Integrin alphaVbeta3; C555023:VPS51 protein, human; D042461:Vascular Endothelial Growth Factor A; D033921:Vesicular Transport Proteins; D042810:Receptor, TIE-1; D042787:Receptor, TIE-2
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