Selective Mitogen Activated Protein Kinase Activity Sensors through the Application of Directionally Programmable D Domain Motifs
Accurate and quantitative methods for measuring the dynamic fluctuations of protein kinase activities are critically needed as diagnostic tools and for the evaluation of kinase-targeted inhibitors, which represent a major therapeutic development area in the treatment of cancer and other diseases. In...
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| Format: | Article |
| Language: | en_US |
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American Chemical Society (ACS)
2016
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| Online Access: | http://hdl.handle.net/1721.1/105102 https://orcid.org/0000-0002-9547-3251 https://orcid.org/0000-0002-5749-7869 |
| _version_ | 1826207291069693952 |
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| author | Peterson, Laura B Yaffe, Michael B Imperiali, Barbara |
| author2 | Massachusetts Institute of Technology. Department of Biological Engineering |
| author_facet | Massachusetts Institute of Technology. Department of Biological Engineering Peterson, Laura B Yaffe, Michael B Imperiali, Barbara |
| author_sort | Peterson, Laura B |
| collection | MIT |
| description | Accurate and quantitative methods for measuring the dynamic fluctuations of protein kinase activities are critically needed as diagnostic tools and for the evaluation of kinase-targeted inhibitors, which represent a major therapeutic development area in the treatment of cancer and other diseases. In particular, rapid and economical methods that utilize simple instrumentation and provide quantitative data in a high throughput format will have the most impact on basic research in systems biology and medicine. There are over 500 protein kinases in the human kinome. Among these, the mitogen activated protein (MAP) kinases are recognized to be central players in key cellular signaling events and are associated with essential processes including growth, proliferation, differentiation, migration, and apoptosis. The major challenge with MAP kinase sensor development is achieving high selectivity since these kinases rely acutely on secondary interactions distal to the phosphorylation site to impart substrate specificity. Herein we describe the development and application of selective sensors for three MAP kinase subfamilies, ERK1/2, p38α/β, and JNK1/2/3. The new sensors are based on a modular design, which includes a sensing element that exploits a sulfonamido-oxine (Sox) fluorophore for reporting phosphorylation, a recognition and specificity element based on reported docking domain motifs and a variable linker, which can be engineered to optimize the intermodule distance and relative orientation. Following rigorous validation, the capabilities of the new sensors are exemplified through the quantitative analysis of the target MAP kinases in breast cancer progression in a cell culture model, which reveals a strong correlation between p38α/β activity and increased tumorgenicity. |
| first_indexed | 2024-09-23T13:47:21Z |
| format | Article |
| id | mit-1721.1/105102 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T13:47:21Z |
| publishDate | 2016 |
| publisher | American Chemical Society (ACS) |
| record_format | dspace |
| spelling | mit-1721.1/1051022022-09-28T16:12:09Z Selective Mitogen Activated Protein Kinase Activity Sensors through the Application of Directionally Programmable D Domain Motifs Peterson, Laura B Yaffe, Michael B Imperiali, Barbara Massachusetts Institute of Technology. Department of Biological Engineering Massachusetts Institute of Technology. Department of Biology Massachusetts Institute of Technology. Department of Chemistry Koch Institute for Integrative Cancer Research at MIT Peterson, Laura B Yaffe, Michael B Imperiali, Barbara Accurate and quantitative methods for measuring the dynamic fluctuations of protein kinase activities are critically needed as diagnostic tools and for the evaluation of kinase-targeted inhibitors, which represent a major therapeutic development area in the treatment of cancer and other diseases. In particular, rapid and economical methods that utilize simple instrumentation and provide quantitative data in a high throughput format will have the most impact on basic research in systems biology and medicine. There are over 500 protein kinases in the human kinome. Among these, the mitogen activated protein (MAP) kinases are recognized to be central players in key cellular signaling events and are associated with essential processes including growth, proliferation, differentiation, migration, and apoptosis. The major challenge with MAP kinase sensor development is achieving high selectivity since these kinases rely acutely on secondary interactions distal to the phosphorylation site to impart substrate specificity. Herein we describe the development and application of selective sensors for three MAP kinase subfamilies, ERK1/2, p38α/β, and JNK1/2/3. The new sensors are based on a modular design, which includes a sensing element that exploits a sulfonamido-oxine (Sox) fluorophore for reporting phosphorylation, a recognition and specificity element based on reported docking domain motifs and a variable linker, which can be engineered to optimize the intermodule distance and relative orientation. Following rigorous validation, the capabilities of the new sensors are exemplified through the quantitative analysis of the target MAP kinases in breast cancer progression in a cell culture model, which reveals a strong correlation between p38α/β activity and increased tumorgenicity. National Institutes of Health (U.S.) (NIH Ruth L. Kirschstein National Research Service Award (F32 GM102992)) David H. Koch Institute for Integrative Cancer Research at MIT (Koch Institute Bridge Project Grant) National Cancer Institute (U.S.) (Integrated Cancer Biology Program at MIT (NIH U54-CA112967)) National Institutes of Health (U.S.) (NIH GM104047) National Institutes of Health (U.S.) (NIH ES015339) 2016-10-26T19:37:39Z 2016-10-26T19:37:39Z 2014-08 2014-08 Article http://purl.org/eprint/type/JournalArticle 0006-2960 1520-4995 http://hdl.handle.net/1721.1/105102 Peterson, Laura B., Michael B. Yaffe, and Barbara Imperiali. “Selective Mitogen Activated Protein Kinase Activity Sensors through the Application of Directionally Programmable D Domain Motifs.” Biochemistry 53, no. 36 (September 16, 2014): 5771–5778. © 2014 American Chemical Society. https://orcid.org/0000-0002-9547-3251 https://orcid.org/0000-0002-5749-7869 en_US http://dx.doi.org/10.1021/bi500862c Biochemistry Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use. application/pdf American Chemical Society (ACS) ACS |
| spellingShingle | Peterson, Laura B Yaffe, Michael B Imperiali, Barbara Selective Mitogen Activated Protein Kinase Activity Sensors through the Application of Directionally Programmable D Domain Motifs |
| title | Selective Mitogen Activated Protein Kinase Activity Sensors through the Application of Directionally Programmable D Domain Motifs |
| title_full | Selective Mitogen Activated Protein Kinase Activity Sensors through the Application of Directionally Programmable D Domain Motifs |
| title_fullStr | Selective Mitogen Activated Protein Kinase Activity Sensors through the Application of Directionally Programmable D Domain Motifs |
| title_full_unstemmed | Selective Mitogen Activated Protein Kinase Activity Sensors through the Application of Directionally Programmable D Domain Motifs |
| title_short | Selective Mitogen Activated Protein Kinase Activity Sensors through the Application of Directionally Programmable D Domain Motifs |
| title_sort | selective mitogen activated protein kinase activity sensors through the application of directionally programmable d domain motifs |
| url | http://hdl.handle.net/1721.1/105102 https://orcid.org/0000-0002-9547-3251 https://orcid.org/0000-0002-5749-7869 |
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