Evidence for allosteric effects on p53 oligomerization induced by phosphorylation
p53 is a tetrameric protein with a thermodynamically unstable deoxyribonucleic acid (DNA)‐binding domain flanked by intrinsically disordered regulatory domains that control its activity. The unstable and disordered segments of p53 allow high flexibility as it interacts with binding partners and perm...
| Main Authors: | , , , , , , |
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| Other Authors: | |
| Format: | Journal Article |
| Language: | English |
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2019
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| Online Access: | https://hdl.handle.net/10356/89905 http://hdl.handle.net/10220/47748 |
| _version_ | 1811681286265241600 |
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| author | Muller, Petr Chan, Juliana M. Simoncik, Oliver Fojta, Miroslav Lane, David P. Hupp, Ted Vojtesek, Borivoj |
| author2 | School of Chemical and Biomedical Engineering |
| author_facet | School of Chemical and Biomedical Engineering Muller, Petr Chan, Juliana M. Simoncik, Oliver Fojta, Miroslav Lane, David P. Hupp, Ted Vojtesek, Borivoj |
| author_sort | Muller, Petr |
| collection | NTU |
| description | p53 is a tetrameric protein with a thermodynamically unstable deoxyribonucleic acid (DNA)‐binding domain flanked by intrinsically disordered regulatory domains that control its activity. The unstable and disordered segments of p53 allow high flexibility as it interacts with binding partners and permits a rapid on/off switch to control its function. The p53 tetramer can exist in multiple conformational states, any of which can be stabilized by a particular modification. Here, we apply the allostery model to p53 to ask whether evidence can be found that the “activating” C‐terminal phosphorylation of p53 stabilizes a specific conformation of the protein in the absence of DNA. We take advantage of monoclonal antibodies for p53 that measure indirectly the following conformations: unfolded, folded, and tetrameric. A double antibody capture enzyme linked‐immunosorbent assay was used to observe evidence of conformational changes of human p53 upon phosphorylation by casein kinase 2 in vitro. It was demonstrated that oligomerization and stabilization of p53 wild‐type conformation results in differential exposure of conformational epitopes PAb1620, PAb240, and DO12 that indicates a reduction in the “unfolded” conformation and increases in the folded conformation coincide with increases in its oligomerization state. These data highlight that the oligomeric conformation of p53 can be stabilized by an activating enzyme and further highlight the utility of the allostery model when applied to understanding the regulation of unstable and intrinsically disordered proteins. |
| first_indexed | 2024-10-01T03:38:32Z |
| format | Journal Article |
| id | ntu-10356/89905 |
| institution | Nanyang Technological University |
| language | English |
| last_indexed | 2024-10-01T03:38:32Z |
| publishDate | 2019 |
| record_format | dspace |
| spelling | ntu-10356/899052020-03-07T11:35:31Z Evidence for allosteric effects on p53 oligomerization induced by phosphorylation Muller, Petr Chan, Juliana M. Simoncik, Oliver Fojta, Miroslav Lane, David P. Hupp, Ted Vojtesek, Borivoj School of Chemical and Biomedical Engineering DRNTU::Science::Chemistry Protein Oligomerization p53 is a tetrameric protein with a thermodynamically unstable deoxyribonucleic acid (DNA)‐binding domain flanked by intrinsically disordered regulatory domains that control its activity. The unstable and disordered segments of p53 allow high flexibility as it interacts with binding partners and permits a rapid on/off switch to control its function. The p53 tetramer can exist in multiple conformational states, any of which can be stabilized by a particular modification. Here, we apply the allostery model to p53 to ask whether evidence can be found that the “activating” C‐terminal phosphorylation of p53 stabilizes a specific conformation of the protein in the absence of DNA. We take advantage of monoclonal antibodies for p53 that measure indirectly the following conformations: unfolded, folded, and tetrameric. A double antibody capture enzyme linked‐immunosorbent assay was used to observe evidence of conformational changes of human p53 upon phosphorylation by casein kinase 2 in vitro. It was demonstrated that oligomerization and stabilization of p53 wild‐type conformation results in differential exposure of conformational epitopes PAb1620, PAb240, and DO12 that indicates a reduction in the “unfolded” conformation and increases in the folded conformation coincide with increases in its oligomerization state. These data highlight that the oligomeric conformation of p53 can be stabilized by an activating enzyme and further highlight the utility of the allostery model when applied to understanding the regulation of unstable and intrinsically disordered proteins. 2019-03-01T07:45:42Z 2019-12-06T17:36:18Z 2019-03-01T07:45:42Z 2019-12-06T17:36:18Z 2017 Journal Article Muller, P., Chan, J. M., Simoncik, O., Fojta, M., Lane, D. P., Hupp, T., & Vojtesek, B. (2018). Evidence for allosteric effects on p53 oligomerization induced by phosphorylation. Protein Science, 27(2), 523-530. doi:http://dx.doi.org/10.1002/pro.3344 0961-8368 https://hdl.handle.net/10356/89905 http://hdl.handle.net/10220/47748 10.1002/pro.3344 en Protein Science © 2017 The Protein Society. All rights reserved. |
| spellingShingle | DRNTU::Science::Chemistry Protein Oligomerization Muller, Petr Chan, Juliana M. Simoncik, Oliver Fojta, Miroslav Lane, David P. Hupp, Ted Vojtesek, Borivoj Evidence for allosteric effects on p53 oligomerization induced by phosphorylation |
| title | Evidence for allosteric effects on p53 oligomerization induced by phosphorylation |
| title_full | Evidence for allosteric effects on p53 oligomerization induced by phosphorylation |
| title_fullStr | Evidence for allosteric effects on p53 oligomerization induced by phosphorylation |
| title_full_unstemmed | Evidence for allosteric effects on p53 oligomerization induced by phosphorylation |
| title_short | Evidence for allosteric effects on p53 oligomerization induced by phosphorylation |
| title_sort | evidence for allosteric effects on p53 oligomerization induced by phosphorylation |
| topic | DRNTU::Science::Chemistry Protein Oligomerization |
| url | https://hdl.handle.net/10356/89905 http://hdl.handle.net/10220/47748 |
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