Complex Formation between VEGFR2 and the β2-Adrenoceptor

Laura E. Kilpatrick, Diana C. Alcobia, Carl W. White, Chloe J. Peach, Jackie R. Glenn, Kris Zimmerman, Alexander Kondrashov, Kevin D.G. Pfleger, Rachel Friedman Ohana, Matthew B. Robers, Keith V. Wood, Erica K. Sloan, Jeanette Woolard, Stephen J. Hill

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Abstract

Vascular endothelial growth factor (VEGF) is an important mediator of endothelial cell proliferation and angiogenesis via its receptor VEGFR2. A common tumor associated with elevated VEGFR2 signaling is infantile hemangioma that is caused by a rapid proliferation of vascular endothelial cells. The current first-line treatment for infantile hemangioma is the β-adrenoceptor antagonist, propranolol, although its mechanism of action is not understood. Here we have used bioluminescence resonance energy transfer and VEGFR2 genetically tagged with NanoLuc luciferase to demonstrate that oligomeric complexes involving VEGFR2 and the β2-adrenoceptor can be generated in both cell membranes and intracellular endosomes. These complexes are induced by agonist treatment and retain their ability to couple to intracellular signaling proteins. Furthermore, coupling of β2-adrenoceptor to β-arrestin2 is prolonged by VEGFR2 activation. These data suggest that protein-protein interactions between VEGFR2, the β2-adrenoceptor, and β-arrestin2 may provide insight into their roles in health and disease. Kilpatrick et al. have used bioluminescence resonance energy transfer (BRET) and VEGFR2 tagged with NanoLuc luciferase, to demonstrate that oligomeric complexes involving VEGFR2 and β2-adrenoceptors can be generated in both cell membranes and intracellular endosomes. These complexes are agonist sensitive and retain their ability to couple to intracellular signaling proteins.

Original languageEnglish
Pages (from-to)830-841.e9
JournalCell Chemical Biology
Volume26
Issue number6
DOIs
Publication statusPublished - 20 Jun 2019

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Adrenergic Receptors
Intracellular Signaling Peptides and Proteins
Bioluminescence
Endosomes
Energy Transfer
Endothelial cells
Cell membranes
Hemangioma
Luciferases
Energy transfer
Endothelial Cells
Cell Membrane
Cell proliferation
Propranolol
Vascular Endothelial Growth Factor A
Tumors
Proteins
Chemical activation
Cell Proliferation
Health

Cite this

Kilpatrick, L. E., Alcobia, D. C., White, C. W., Peach, C. J., Glenn, J. R., Zimmerman, K., ... Hill, S. J. (2019). Complex Formation between VEGFR2 and the β2-Adrenoceptor. Cell Chemical Biology, 26(6), 830-841.e9. https://doi.org/10.1016/j.chembiol.2019.02.014
Kilpatrick, Laura E. ; Alcobia, Diana C. ; White, Carl W. ; Peach, Chloe J. ; Glenn, Jackie R. ; Zimmerman, Kris ; Kondrashov, Alexander ; Pfleger, Kevin D.G. ; Ohana, Rachel Friedman ; Robers, Matthew B. ; Wood, Keith V. ; Sloan, Erica K. ; Woolard, Jeanette ; Hill, Stephen J. / Complex Formation between VEGFR2 and the β2-Adrenoceptor. In: Cell Chemical Biology. 2019 ; Vol. 26, No. 6. pp. 830-841.e9.
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Kilpatrick, LE, Alcobia, DC, White, CW, Peach, CJ, Glenn, JR, Zimmerman, K, Kondrashov, A, Pfleger, KDG, Ohana, RF, Robers, MB, Wood, KV, Sloan, EK, Woolard, J & Hill, SJ 2019, 'Complex Formation between VEGFR2 and the β2-Adrenoceptor' Cell Chemical Biology, vol. 26, no. 6, pp. 830-841.e9. https://doi.org/10.1016/j.chembiol.2019.02.014

Complex Formation between VEGFR2 and the β2-Adrenoceptor. / Kilpatrick, Laura E.; Alcobia, Diana C.; White, Carl W.; Peach, Chloe J.; Glenn, Jackie R.; Zimmerman, Kris; Kondrashov, Alexander; Pfleger, Kevin D.G.; Ohana, Rachel Friedman; Robers, Matthew B.; Wood, Keith V.; Sloan, Erica K.; Woolard, Jeanette; Hill, Stephen J.

In: Cell Chemical Biology, Vol. 26, No. 6, 20.06.2019, p. 830-841.e9.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Complex Formation between VEGFR2 and the β2-Adrenoceptor

AU - Kilpatrick, Laura E.

AU - Alcobia, Diana C.

AU - White, Carl W.

AU - Peach, Chloe J.

AU - Glenn, Jackie R.

AU - Zimmerman, Kris

AU - Kondrashov, Alexander

AU - Pfleger, Kevin D.G.

AU - Ohana, Rachel Friedman

AU - Robers, Matthew B.

AU - Wood, Keith V.

AU - Sloan, Erica K.

AU - Woolard, Jeanette

AU - Hill, Stephen J.

PY - 2019/6/20

Y1 - 2019/6/20

N2 - Vascular endothelial growth factor (VEGF) is an important mediator of endothelial cell proliferation and angiogenesis via its receptor VEGFR2. A common tumor associated with elevated VEGFR2 signaling is infantile hemangioma that is caused by a rapid proliferation of vascular endothelial cells. The current first-line treatment for infantile hemangioma is the β-adrenoceptor antagonist, propranolol, although its mechanism of action is not understood. Here we have used bioluminescence resonance energy transfer and VEGFR2 genetically tagged with NanoLuc luciferase to demonstrate that oligomeric complexes involving VEGFR2 and the β2-adrenoceptor can be generated in both cell membranes and intracellular endosomes. These complexes are induced by agonist treatment and retain their ability to couple to intracellular signaling proteins. Furthermore, coupling of β2-adrenoceptor to β-arrestin2 is prolonged by VEGFR2 activation. These data suggest that protein-protein interactions between VEGFR2, the β2-adrenoceptor, and β-arrestin2 may provide insight into their roles in health and disease. Kilpatrick et al. have used bioluminescence resonance energy transfer (BRET) and VEGFR2 tagged with NanoLuc luciferase, to demonstrate that oligomeric complexes involving VEGFR2 and β2-adrenoceptors can be generated in both cell membranes and intracellular endosomes. These complexes are agonist sensitive and retain their ability to couple to intracellular signaling proteins.

AB - Vascular endothelial growth factor (VEGF) is an important mediator of endothelial cell proliferation and angiogenesis via its receptor VEGFR2. A common tumor associated with elevated VEGFR2 signaling is infantile hemangioma that is caused by a rapid proliferation of vascular endothelial cells. The current first-line treatment for infantile hemangioma is the β-adrenoceptor antagonist, propranolol, although its mechanism of action is not understood. Here we have used bioluminescence resonance energy transfer and VEGFR2 genetically tagged with NanoLuc luciferase to demonstrate that oligomeric complexes involving VEGFR2 and the β2-adrenoceptor can be generated in both cell membranes and intracellular endosomes. These complexes are induced by agonist treatment and retain their ability to couple to intracellular signaling proteins. Furthermore, coupling of β2-adrenoceptor to β-arrestin2 is prolonged by VEGFR2 activation. These data suggest that protein-protein interactions between VEGFR2, the β2-adrenoceptor, and β-arrestin2 may provide insight into their roles in health and disease. Kilpatrick et al. have used bioluminescence resonance energy transfer (BRET) and VEGFR2 tagged with NanoLuc luciferase, to demonstrate that oligomeric complexes involving VEGFR2 and β2-adrenoceptors can be generated in both cell membranes and intracellular endosomes. These complexes are agonist sensitive and retain their ability to couple to intracellular signaling proteins.

KW - BRET

KW - CRISPR/Cas9

KW - infantile haemangioma

KW - NanoBRET

KW - receptor oligomerisation

KW - VEGFR2

KW - β-arrestin

KW - β-adrenoceptors

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U2 - 10.1016/j.chembiol.2019.02.014

DO - 10.1016/j.chembiol.2019.02.014

M3 - Article

VL - 26

SP - 830-841.e9

JO - Cell Chemical Biology

JF - Cell Chemical Biology

SN - 2451-9456

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ER -

Kilpatrick LE, Alcobia DC, White CW, Peach CJ, Glenn JR, Zimmerman K et al. Complex Formation between VEGFR2 and the β2-Adrenoceptor. Cell Chemical Biology. 2019 Jun 20;26(6):830-841.e9. https://doi.org/10.1016/j.chembiol.2019.02.014