Fluorescence Correlation Spectroscopy in Drug Discovery: Study of Alexa532-Endothelin 1 Binding to the Endothelin ETA Receptor to Describe the Pharmacological Profile of Natural Products
Fluorescence correlation spectroscopy and the newly synthesized Alexa532-ET1 were used to study the dynamics of the endothelin ETA receptor-ligand complex alone and under the influence of a semisynthetic selective antagonist and a fungal extract on living A10 cells. Dose-dependent increase of inosit...
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| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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Wiley
2012-01-01
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| Series: | The Scientific World Journal |
| Online Access: | http://dx.doi.org/10.1100/2012/524169 |
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| author | Catherina Caballero-George Thomas Sorkalla Daniel Jakobs Jessica Bolaños Huzefa Raja Carol Shearer Eldredge Bermingham Hanns Häberlein |
| author_facet | Catherina Caballero-George Thomas Sorkalla Daniel Jakobs Jessica Bolaños Huzefa Raja Carol Shearer Eldredge Bermingham Hanns Häberlein |
| author_sort | Catherina Caballero-George |
| collection | DOAJ |
| description | Fluorescence correlation spectroscopy and the newly synthesized Alexa532-ET1 were used to study the dynamics of the endothelin ETA receptor-ligand complex alone and under the influence of a semisynthetic selective antagonist and a fungal extract on living A10 cells. Dose-dependent increase of inositol phosphate production was seen for Alexa532-ET1, and its binding was reduced to 8% by the selective endothelin ETA antagonist BQ-123, confirming the specific binding of Alexa532-ET1 to the endothelin ETA receptor. Two different lateral mobilities of the receptor-ligand complexes within the cell membrane were found allowing the discrimination of different states for this complex. BQ-123 showed a strong binding affinity to the “inactive” receptor state characterized by the slow diffusion time constant. A similar effect was observed for the fungal extract, which completely displaced Alexa532-ET1 from its binding to the “inactive” receptor state. These findings suggest that both BQ-123 and the fungal extract act as inverse agonists. |
| format | Article |
| id | doaj-art-48243669ae214fbb8655d5103b7bc229 |
| institution | Kabale University |
| issn | 1537-744X |
| language | English |
| publishDate | 2012-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | The Scientific World Journal |
| spelling | doaj-art-48243669ae214fbb8655d5103b7bc2292025-02-03T05:43:54ZengWileyThe Scientific World Journal1537-744X2012-01-01201210.1100/2012/524169524169Fluorescence Correlation Spectroscopy in Drug Discovery: Study of Alexa532-Endothelin 1 Binding to the Endothelin ETA Receptor to Describe the Pharmacological Profile of Natural ProductsCatherina Caballero-George0Thomas Sorkalla1Daniel Jakobs2Jessica Bolaños3Huzefa Raja4Carol Shearer5Eldredge Bermingham6Hanns Häberlein7Department of Molecular Pharmacology and Pharmacognosy, Drug Discovery Center, Institute for Scientific Research and High Technology Services, P.O. Box 0843-01103, Panama, PanamaInstitute of Biochemistry and Molecular Biology, Rheinische Friedrich-Wilhelms-University, Nussallee 11, 53115 Bonn, GermanyInstitute of Biochemistry and Molecular Biology, Rheinische Friedrich-Wilhelms-University, Nussallee 11, 53115 Bonn, GermanyDepartment of Molecular Pharmacology and Pharmacognosy, Drug Discovery Center, Institute for Scientific Research and High Technology Services, P.O. Box 0843-01103, Panama, PanamaDepartment of Plant Biology, University of Illinois at Urbana-Champaign, Room 265, Morrill Hall, 505 S. Goodwin Avenue, Urbana, IL 61801, USADepartment of Plant Biology, University of Illinois at Urbana-Champaign, Room 265, Morrill Hall, 505 S. Goodwin Avenue, Urbana, IL 61801, USASmithsonian Tropical Research Institute, P.O. Box 0843-03092, Panama, PanamaInstitute of Biochemistry and Molecular Biology, Rheinische Friedrich-Wilhelms-University, Nussallee 11, 53115 Bonn, GermanyFluorescence correlation spectroscopy and the newly synthesized Alexa532-ET1 were used to study the dynamics of the endothelin ETA receptor-ligand complex alone and under the influence of a semisynthetic selective antagonist and a fungal extract on living A10 cells. Dose-dependent increase of inositol phosphate production was seen for Alexa532-ET1, and its binding was reduced to 8% by the selective endothelin ETA antagonist BQ-123, confirming the specific binding of Alexa532-ET1 to the endothelin ETA receptor. Two different lateral mobilities of the receptor-ligand complexes within the cell membrane were found allowing the discrimination of different states for this complex. BQ-123 showed a strong binding affinity to the “inactive” receptor state characterized by the slow diffusion time constant. A similar effect was observed for the fungal extract, which completely displaced Alexa532-ET1 from its binding to the “inactive” receptor state. These findings suggest that both BQ-123 and the fungal extract act as inverse agonists.http://dx.doi.org/10.1100/2012/524169 |
| spellingShingle | Catherina Caballero-George Thomas Sorkalla Daniel Jakobs Jessica Bolaños Huzefa Raja Carol Shearer Eldredge Bermingham Hanns Häberlein Fluorescence Correlation Spectroscopy in Drug Discovery: Study of Alexa532-Endothelin 1 Binding to the Endothelin ETA Receptor to Describe the Pharmacological Profile of Natural Products The Scientific World Journal |
| title | Fluorescence Correlation Spectroscopy in Drug Discovery: Study of Alexa532-Endothelin 1 Binding to the Endothelin ETA Receptor to Describe the Pharmacological Profile of Natural Products |
| title_full | Fluorescence Correlation Spectroscopy in Drug Discovery: Study of Alexa532-Endothelin 1 Binding to the Endothelin ETA Receptor to Describe the Pharmacological Profile of Natural Products |
| title_fullStr | Fluorescence Correlation Spectroscopy in Drug Discovery: Study of Alexa532-Endothelin 1 Binding to the Endothelin ETA Receptor to Describe the Pharmacological Profile of Natural Products |
| title_full_unstemmed | Fluorescence Correlation Spectroscopy in Drug Discovery: Study of Alexa532-Endothelin 1 Binding to the Endothelin ETA Receptor to Describe the Pharmacological Profile of Natural Products |
| title_short | Fluorescence Correlation Spectroscopy in Drug Discovery: Study of Alexa532-Endothelin 1 Binding to the Endothelin ETA Receptor to Describe the Pharmacological Profile of Natural Products |
| title_sort | fluorescence correlation spectroscopy in drug discovery study of alexa532 endothelin 1 binding to the endothelin eta receptor to describe the pharmacological profile of natural products |
| url | http://dx.doi.org/10.1100/2012/524169 |
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