The membrane-permeable calmodulin inhibitors (W-7/W-13) bind to membranes changing the electrostatic surface potential. Dual effect of W-13 on EGFR activation
Membrane-permeable calmodulin inhibitors, such as the napthalenesulfonamide derivatives W-7/W-13, trifluoperazine, and calmidazolium, are used widely to investigate the role of calcium/calmodulin (Ca2+/CaM) in living cells. If two chemically different inhibitors (e.g. W-7 and trifluoperazine) produc...
| Authors: | ; ; ; ; ; ; ; |
|---|---|
| Format: | Article |
| Status: | Accepted version |
| Publication date: | 2007 |
| Country: | España |
| Institution: | Universidad de Barcelona |
| Repository: | Dipòsit Digital de la UB |
| OAI Identifier: | oai:diposit.ub.edu:2445/33849 |
| Online Access: | https://hdl.handle.net/2445/33849 |
| Access Level: | Open access |
| Keyword: | Calmodulina Factor de creixement epidèrmic Calmodulin Epidermal growth factor |
| _version_ | 1878731332126769152 |
|---|---|
| author | Sengupta, Parijat |
| author2 | Ruano, María José Tebar Ramon, Francesc Golebiewska, Urszula Zaitseva, Irina Enrich Bastús, Carles McLaughlin, Stuart Villalobo, Antonio |
| author2_role | author author author author author author author |
| author_browse | Enrich Bastús, Carles Golebiewska, Urszula McLaughlin, Stuart Ruano, María José Sengupta, Parijat Tebar Ramon, Francesc Villalobo, Antonio Zaitseva, Irina |
| author_facet | Sengupta, Parijat Ruano, María José Tebar Ramon, Francesc Golebiewska, Urszula Zaitseva, Irina Enrich Bastús, Carles McLaughlin, Stuart Villalobo, Antonio |
| author_role | author |
| collection | Dipòsit Digital de la UB |
| description | Membrane-permeable calmodulin inhibitors, such as the napthalenesulfonamide derivatives W-7/W-13, trifluoperazine, and calmidazolium, are used widely to investigate the role of calcium/calmodulin (Ca2+/CaM) in living cells. If two chemically different inhibitors (e.g. W-7 and trifluoperazine) produce similar effects, investigators often assume the effects are due to CaM inhibition. Zeta potential measurements, however, show that these amphipathic weak bases bind to phospholipid vesicles at the same concentrations as they inhibit Ca 2 /CaM; this suggests that they also bind to the inner leaflet of the plasma membrane, reducing its negative electrostatic surface potential. This change will cause electrostatically bound clusters of basic residues on peripheral (e.g. Src and K-Ras4B) and integral (e.g. epidermal growth factor receptor (EGFR)) proteins to translocate from the membrane to the cytoplasm. We measured inhibitor-mediated translocation of a simple basic peptide corresponding to the calmodulin-binding juxtamembrane region of the EGFR on model membranes; W-7/W-13 causes translocation of this peptide from membrane to solution, suggesting that caution must be exercised when interpreting the results obtained with these inhibitors in living cells. We present evidence that they exert dual effects on autophosphorylation of EGFR;W-13 inhibits epidermal growth factordependent EGFR autophosphorylation under different experimental conditions, but in the absence of epidermal growth factor, W-13 stimulates autophosphorylation of the receptor in four different cell types. Our interpretation is that the former effect is due toW-13inhibition of Ca 2 /CaM, but thelatter results could be due to binding of W-13 to the plasma membrane. |
| eu_rights_str_mv | openAccess |
| format | article |
| id | ES_2d776cdef8d397f09efbac4f044d1ebc |
| institution | Universidad de Barcelona |
| instname_str | Universidad de Barcelona |
| network_acronym_str | ES |
| network_name_str | España |
| oai_identifier_str | oai:diposit.ub.edu:2445/33849 |
| publishDate | 2007 |
| publishDateSort | 2007 |
| publisher | American Society for Biochemistry and Molecular Biology |
| publisherStr | American Society for Biochemistry and Molecular Biology |
| reponame_str | Dipòsit Digital de la UB |
| spelling | The membrane-permeable calmodulin inhibitors (W-7/W-13) bind to membranes changing the electrostatic surface potential. Dual effect of W-13 on EGFR activation Sengupta, Parijat Ruano, María José Tebar Ramon, Francesc Golebiewska, Urszula Zaitseva, Irina Enrich Bastús, Carles McLaughlin, Stuart Villalobo, Antonio Calmodulina Factor de creixement epidèrmic Calmodulin Epidermal growth factor Membrane-permeable calmodulin inhibitors, such as the napthalenesulfonamide derivatives W-7/W-13, trifluoperazine, and calmidazolium, are used widely to investigate the role of calcium/calmodulin (Ca2+/CaM) in living cells. If two chemically different inhibitors (e.g. W-7 and trifluoperazine) produce similar effects, investigators often assume the effects are due to CaM inhibition. Zeta potential measurements, however, show that these amphipathic weak bases bind to phospholipid vesicles at the same concentrations as they inhibit Ca 2 /CaM; this suggests that they also bind to the inner leaflet of the plasma membrane, reducing its negative electrostatic surface potential. This change will cause electrostatically bound clusters of basic residues on peripheral (e.g. Src and K-Ras4B) and integral (e.g. epidermal growth factor receptor (EGFR)) proteins to translocate from the membrane to the cytoplasm. We measured inhibitor-mediated translocation of a simple basic peptide corresponding to the calmodulin-binding juxtamembrane region of the EGFR on model membranes; W-7/W-13 causes translocation of this peptide from membrane to solution, suggesting that caution must be exercised when interpreting the results obtained with these inhibitors in living cells. We present evidence that they exert dual effects on autophosphorylation of EGFR;W-13 inhibits epidermal growth factordependent EGFR autophosphorylation under different experimental conditions, but in the absence of epidermal growth factor, W-13 stimulates autophosphorylation of the receptor in four different cell types. Our interpretation is that the former effect is due toW-13inhibition of Ca 2 /CaM, but thelatter results could be due to binding of W-13 to the plasma membrane. American Society for Biochemistry and Molecular Biology https://hdl.handle.net/2445/33849 |
| spellingShingle | The membrane-permeable calmodulin inhibitors (W-7/W-13) bind to membranes changing the electrostatic surface potential. Dual effect of W-13 on EGFR activation Sengupta, Parijat Calmodulina Factor de creixement epidèrmic Calmodulin Epidermal growth factor |
| status_str | acceptedVersion |
| title | The membrane-permeable calmodulin inhibitors (W-7/W-13) bind to membranes changing the electrostatic surface potential. Dual effect of W-13 on EGFR activation |
| title_full | The membrane-permeable calmodulin inhibitors (W-7/W-13) bind to membranes changing the electrostatic surface potential. Dual effect of W-13 on EGFR activation |
| title_fullStr | The membrane-permeable calmodulin inhibitors (W-7/W-13) bind to membranes changing the electrostatic surface potential. Dual effect of W-13 on EGFR activation |
| title_full_unstemmed | The membrane-permeable calmodulin inhibitors (W-7/W-13) bind to membranes changing the electrostatic surface potential. Dual effect of W-13 on EGFR activation |
| title_short | The membrane-permeable calmodulin inhibitors (W-7/W-13) bind to membranes changing the electrostatic surface potential. Dual effect of W-13 on EGFR activation |
| title_sort | The membrane-permeable calmodulin inhibitors (W-7/W-13) bind to membranes changing the electrostatic surface potential. Dual effect of W-13 on EGFR activation |
| topic | Calmodulina Factor de creixement epidèrmic Calmodulin Epidermal growth factor |
| topic_facet | Calmodulina Factor de creixement epidèrmic Calmodulin Epidermal growth factor |
| url | https://hdl.handle.net/2445/33849 |