Chemical Inhibitors of Dynamin Exert Differential Effects in VEGF Signaling
VEGFR2 is the main receptor and mediator of the vasculogenic and angiogenic activity of VEGF. Activated VEGFR2 internalizes through clathrin-mediated endocytosis and macropinocytosis. As dynamin is a key regulator of the clathrin pathway, chemical inhibitors of dynamin are commonly used to assess th...
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MDPI AG
2021-04-01
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| author | Dimitris Basagiannis Sofia Zografou Evangeli Goula Despoina Gkeka Evangelos Kolettas Savvas Christoforidis |
| author_facet | Dimitris Basagiannis Sofia Zografou Evangeli Goula Despoina Gkeka Evangelos Kolettas Savvas Christoforidis |
| author_sort | Dimitris Basagiannis |
| collection | DOAJ |
| description | VEGFR2 is the main receptor and mediator of the vasculogenic and angiogenic activity of VEGF. Activated VEGFR2 internalizes through clathrin-mediated endocytosis and macropinocytosis. As dynamin is a key regulator of the clathrin pathway, chemical inhibitors of dynamin are commonly used to assess the role of the clathrin route in receptor signaling. However, drugs may also exert off-target effects. Here, we compare the effects of three dynamin inhibitors, dynasore, dyngo 4a and dynole, on VEGFR2 internalization and signaling. Although these drugs consistently inhibit clathrin-mediated endocytosis of both transferrin (a typical cargo of this route) and VEGFR2, surprisingly, they exert contradictory effects in receptor signaling. Thus, while dynasore has no effect on phosphorylation of VEGFR2, the other two drugs are strong inhibitors. Furthermore, although dyngo does not interfere with phosphorylation of Akt, dynasore and dynole have a strong inhibitory effect. These inconsistent effects suggest that the above dynamin blockers, besides inhibiting dynamin-dependent endocytosis of VEGFR2, exert additional inhibitory effects on signaling that are independent of endocytosis; i.e., they are due to off-target effects. Using a recently developed protocol, we comparatively validate the specificity of two endocytic inhibitors, dynasore and EIPA. Our findings highlight the importance of assessing whether the effect of an endocytic drug on signaling is specifically due to its interference with endocytosis or due to off-targets. |
| first_indexed | 2024-03-10T12:02:01Z |
| format | Article |
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| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-10T12:02:01Z |
| publishDate | 2021-04-01 |
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| series | Cells |
| spelling | doaj.art-93ca219adfce4200bcdfb2ebce8676f22023-11-21T16:55:24ZengMDPI AGCells2073-44092021-04-0110599710.3390/cells10050997Chemical Inhibitors of Dynamin Exert Differential Effects in VEGF SignalingDimitris Basagiannis0Sofia Zografou1Evangeli Goula2Despoina Gkeka3Evangelos Kolettas4Savvas Christoforidis5Department of Biomedical Research, Foundation for Research and Technology, Institute of Molecular Biology and Biotechnology, 45110 Ioannina, GreeceDepartment of Biomedical Research, Foundation for Research and Technology, Institute of Molecular Biology and Biotechnology, 45110 Ioannina, GreeceDepartment of Biomedical Research, Foundation for Research and Technology, Institute of Molecular Biology and Biotechnology, 45110 Ioannina, GreeceDepartment of Biomedical Research, Foundation for Research and Technology, Institute of Molecular Biology and Biotechnology, 45110 Ioannina, GreeceDepartment of Biomedical Research, Foundation for Research and Technology, Institute of Molecular Biology and Biotechnology, 45110 Ioannina, GreeceDepartment of Biomedical Research, Foundation for Research and Technology, Institute of Molecular Biology and Biotechnology, 45110 Ioannina, GreeceVEGFR2 is the main receptor and mediator of the vasculogenic and angiogenic activity of VEGF. Activated VEGFR2 internalizes through clathrin-mediated endocytosis and macropinocytosis. As dynamin is a key regulator of the clathrin pathway, chemical inhibitors of dynamin are commonly used to assess the role of the clathrin route in receptor signaling. However, drugs may also exert off-target effects. Here, we compare the effects of three dynamin inhibitors, dynasore, dyngo 4a and dynole, on VEGFR2 internalization and signaling. Although these drugs consistently inhibit clathrin-mediated endocytosis of both transferrin (a typical cargo of this route) and VEGFR2, surprisingly, they exert contradictory effects in receptor signaling. Thus, while dynasore has no effect on phosphorylation of VEGFR2, the other two drugs are strong inhibitors. Furthermore, although dyngo does not interfere with phosphorylation of Akt, dynasore and dynole have a strong inhibitory effect. These inconsistent effects suggest that the above dynamin blockers, besides inhibiting dynamin-dependent endocytosis of VEGFR2, exert additional inhibitory effects on signaling that are independent of endocytosis; i.e., they are due to off-target effects. Using a recently developed protocol, we comparatively validate the specificity of two endocytic inhibitors, dynasore and EIPA. Our findings highlight the importance of assessing whether the effect of an endocytic drug on signaling is specifically due to its interference with endocytosis or due to off-targets.https://www.mdpi.com/2073-4409/10/5/997cell signalingendocytosisvesicular traffickingdynamininhibitorsendothelial cell biology |
| spellingShingle | Dimitris Basagiannis Sofia Zografou Evangeli Goula Despoina Gkeka Evangelos Kolettas Savvas Christoforidis Chemical Inhibitors of Dynamin Exert Differential Effects in VEGF Signaling Cells cell signaling endocytosis vesicular trafficking dynamin inhibitors endothelial cell biology |
| title | Chemical Inhibitors of Dynamin Exert Differential Effects in VEGF Signaling |
| title_full | Chemical Inhibitors of Dynamin Exert Differential Effects in VEGF Signaling |
| title_fullStr | Chemical Inhibitors of Dynamin Exert Differential Effects in VEGF Signaling |
| title_full_unstemmed | Chemical Inhibitors of Dynamin Exert Differential Effects in VEGF Signaling |
| title_short | Chemical Inhibitors of Dynamin Exert Differential Effects in VEGF Signaling |
| title_sort | chemical inhibitors of dynamin exert differential effects in vegf signaling |
| topic | cell signaling endocytosis vesicular trafficking dynamin inhibitors endothelial cell biology |
| url | https://www.mdpi.com/2073-4409/10/5/997 |
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