BRET-based RAS biosensors that show a novel small molecule is an inhibitor of RAS-effector protein-protein interactions
The RAS family of proteins is amongst the most highly mutated in human cancers and has so far eluded drug therapy. Currently, much effort is being made to discover mutant RAS inhibitors and in vitro screening for RAS-binding drugs must be followed by cell-based assays. Here, we have developed a robu...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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eLife Sciences Publications Ltd
2018-07-01
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| Series: | eLife |
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| Online Access: | https://elifesciences.org/articles/37122 |
| _version_ | 1811253374287347712 |
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| author | Nicolas Bery Abimael Cruz-Migoni Carole JR Bataille Camilo E Quevedo Hanna Tulmin Ami Miller Angela Russell Simon EV Phillips Stephen B Carr Terence H Rabbitts |
| author_facet | Nicolas Bery Abimael Cruz-Migoni Carole JR Bataille Camilo E Quevedo Hanna Tulmin Ami Miller Angela Russell Simon EV Phillips Stephen B Carr Terence H Rabbitts |
| author_sort | Nicolas Bery |
| collection | DOAJ |
| description | The RAS family of proteins is amongst the most highly mutated in human cancers and has so far eluded drug therapy. Currently, much effort is being made to discover mutant RAS inhibitors and in vitro screening for RAS-binding drugs must be followed by cell-based assays. Here, we have developed a robust set of bioluminescence resonance energy transfer (BRET)-based RAS biosensors that enable monitoring of RAS-effector interaction inhibition in living cells. These include KRAS, HRAS and NRAS and a variety of different mutations that mirror those found in human cancers with the major RAS effectors such as CRAF, PI3K and RALGDS. We highlighted the utility of these RAS biosensors by showing a RAS-binding compound is a potent pan-RAS-effector interactions inhibitor in cells. The RAS biosensors represent a useful tool to investigate and characterize the potency of anti-RAS inhibitors in cells and more generally any RAS protein-protein interaction (PPI) in cells. |
| first_indexed | 2024-04-12T16:49:57Z |
| format | Article |
| id | doaj.art-6d8a1e8ab8724fc883df0eb1fdb7cab0 |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-12T16:49:57Z |
| publishDate | 2018-07-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-6d8a1e8ab8724fc883df0eb1fdb7cab02022-12-22T03:24:26ZengeLife Sciences Publications LtdeLife2050-084X2018-07-01710.7554/eLife.37122BRET-based RAS biosensors that show a novel small molecule is an inhibitor of RAS-effector protein-protein interactionsNicolas Bery0https://orcid.org/0000-0002-2643-3897Abimael Cruz-Migoni1Carole JR Bataille2Camilo E Quevedo3Hanna Tulmin4Ami Miller5Angela Russell6Simon EV Phillips7Stephen B Carr8Terence H Rabbitts9https://orcid.org/0000-0002-4982-2609MRC Molecular Haematology Unit, Weatherall Institute of Molecular Medicine, University of Oxford, Oxford, United KingdomMRC Molecular Haematology Unit, Weatherall Institute of Molecular Medicine, University of Oxford, Oxford, United Kingdom; Research Complex at Harwell, Rutherford Appleton Laboratory, Didcot, United KingdomChemistry Research Laboratory, Oxford, United KingdomMRC Molecular Haematology Unit, Weatherall Institute of Molecular Medicine, University of Oxford, Oxford, United KingdomMRC Molecular Haematology Unit, Weatherall Institute of Molecular Medicine, University of Oxford, Oxford, United KingdomMRC Molecular Haematology Unit, Weatherall Institute of Molecular Medicine, University of Oxford, Oxford, United KingdomChemistry Research Laboratory, Oxford, United KingdomDepartment of Biochemistry, University of Oxford, Oxford, United KingdomResearch Complex at Harwell, Rutherford Appleton Laboratory, Didcot, United Kingdom; Department of Biochemistry, University of Oxford, Oxford, United KingdomMRC Molecular Haematology Unit, Weatherall Institute of Molecular Medicine, University of Oxford, Oxford, United KingdomThe RAS family of proteins is amongst the most highly mutated in human cancers and has so far eluded drug therapy. Currently, much effort is being made to discover mutant RAS inhibitors and in vitro screening for RAS-binding drugs must be followed by cell-based assays. Here, we have developed a robust set of bioluminescence resonance energy transfer (BRET)-based RAS biosensors that enable monitoring of RAS-effector interaction inhibition in living cells. These include KRAS, HRAS and NRAS and a variety of different mutations that mirror those found in human cancers with the major RAS effectors such as CRAF, PI3K and RALGDS. We highlighted the utility of these RAS biosensors by showing a RAS-binding compound is a potent pan-RAS-effector interactions inhibitor in cells. The RAS biosensors represent a useful tool to investigate and characterize the potency of anti-RAS inhibitors in cells and more generally any RAS protein-protein interaction (PPI) in cells.https://elifesciences.org/articles/37122RASBRETbiosensorsprotein-protein interaction inhibitioncell-based assaysintracellular antibody |
| spellingShingle | Nicolas Bery Abimael Cruz-Migoni Carole JR Bataille Camilo E Quevedo Hanna Tulmin Ami Miller Angela Russell Simon EV Phillips Stephen B Carr Terence H Rabbitts BRET-based RAS biosensors that show a novel small molecule is an inhibitor of RAS-effector protein-protein interactions eLife RAS BRET biosensors protein-protein interaction inhibition cell-based assays intracellular antibody |
| title | BRET-based RAS biosensors that show a novel small molecule is an inhibitor of RAS-effector protein-protein interactions |
| title_full | BRET-based RAS biosensors that show a novel small molecule is an inhibitor of RAS-effector protein-protein interactions |
| title_fullStr | BRET-based RAS biosensors that show a novel small molecule is an inhibitor of RAS-effector protein-protein interactions |
| title_full_unstemmed | BRET-based RAS biosensors that show a novel small molecule is an inhibitor of RAS-effector protein-protein interactions |
| title_short | BRET-based RAS biosensors that show a novel small molecule is an inhibitor of RAS-effector protein-protein interactions |
| title_sort | bret based ras biosensors that show a novel small molecule is an inhibitor of ras effector protein protein interactions |
| topic | RAS BRET biosensors protein-protein interaction inhibition cell-based assays intracellular antibody |
| url | https://elifesciences.org/articles/37122 |
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