Light-regulated allosteric switch enables temporal and subcellular control of enzyme activity
© 2020, eLife Sciences Publications Ltd. All rights reserved. Engineered allosteric regulation of protein activity provides significant advantages for the development of robust and broadly applicable tools. However, the application of allosteric switches in optogenetics has been scarce and suffers f...
| Main Authors: | , , , , , , , , , , , , , , |
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| Other Authors: | |
| Format: | Article |
| Language: | English |
| Published: |
eLife Sciences Publications, Ltd
2021
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| Online Access: | https://hdl.handle.net/1721.1/133606 |
| _version_ | 1826211715832872960 |
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| author | Shaaya, Mark Fauser, Jordan Zhurikhina, Anastasia Conage-Pough, Jason E Huyot, Vincent Brennan, Martin Flower, Cameron T Matsche, Jacob Khan, Shahzeb Natarajan, Viswanathan Rehman, Jalees Kota, Pradeep White, Forest M Tsygankov, Denis Karginov, Andrei V |
| author2 | Koch Institute for Integrative Cancer Research at MIT |
| author_facet | Koch Institute for Integrative Cancer Research at MIT Shaaya, Mark Fauser, Jordan Zhurikhina, Anastasia Conage-Pough, Jason E Huyot, Vincent Brennan, Martin Flower, Cameron T Matsche, Jacob Khan, Shahzeb Natarajan, Viswanathan Rehman, Jalees Kota, Pradeep White, Forest M Tsygankov, Denis Karginov, Andrei V |
| author_sort | Shaaya, Mark |
| collection | MIT |
| description | © 2020, eLife Sciences Publications Ltd. All rights reserved. Engineered allosteric regulation of protein activity provides significant advantages for the development of robust and broadly applicable tools. However, the application of allosteric switches in optogenetics has been scarce and suffers from critical limitations. Here, we report an optogenetic approach that utilizes an engineered Light-Regulated (LightR) allosteric switch module to achieve tight spatiotemporal control of enzymatic activity. Using the tyrosine kinase Src as a model, we demonstrate efficient regulation of the kinase and identify temporally distinct signaling responses ranging from seconds to minutes. LightR-Src off-kinetics can be tuned by modulating the LightR photoconversion cycle. A fast cycling variant enables the stimulation of transient pulses and local regulation of activity in a selected region of a cell. The design of the LightR module ensures broad applicability of the tool, as we demonstrate by achieving light-mediated regulation of Abl and bRaf kinases as well as Cre recombinase. |
| first_indexed | 2024-09-23T15:10:23Z |
| format | Article |
| id | mit-1721.1/133606 |
| institution | Massachusetts Institute of Technology |
| language | English |
| last_indexed | 2024-09-23T15:10:23Z |
| publishDate | 2021 |
| publisher | eLife Sciences Publications, Ltd |
| record_format | dspace |
| spelling | mit-1721.1/1336062023-12-08T17:36:27Z Light-regulated allosteric switch enables temporal and subcellular control of enzyme activity Shaaya, Mark Fauser, Jordan Zhurikhina, Anastasia Conage-Pough, Jason E Huyot, Vincent Brennan, Martin Flower, Cameron T Matsche, Jacob Khan, Shahzeb Natarajan, Viswanathan Rehman, Jalees Kota, Pradeep White, Forest M Tsygankov, Denis Karginov, Andrei V Koch Institute for Integrative Cancer Research at MIT Center for Precision Cancer Medicine Massachusetts Institute of Technology. Department of Biological Engineering Massachusetts Institute of Technology. Computational and Systems Biology Program © 2020, eLife Sciences Publications Ltd. All rights reserved. Engineered allosteric regulation of protein activity provides significant advantages for the development of robust and broadly applicable tools. However, the application of allosteric switches in optogenetics has been scarce and suffers from critical limitations. Here, we report an optogenetic approach that utilizes an engineered Light-Regulated (LightR) allosteric switch module to achieve tight spatiotemporal control of enzymatic activity. Using the tyrosine kinase Src as a model, we demonstrate efficient regulation of the kinase and identify temporally distinct signaling responses ranging from seconds to minutes. LightR-Src off-kinetics can be tuned by modulating the LightR photoconversion cycle. A fast cycling variant enables the stimulation of transient pulses and local regulation of activity in a selected region of a cell. The design of the LightR module ensures broad applicability of the tool, as we demonstrate by achieving light-mediated regulation of Abl and bRaf kinases as well as Cre recombinase. 2021-10-27T19:53:47Z 2021-10-27T19:53:47Z 2020 2021-09-10T17:42:51Z Article http://purl.org/eprint/type/JournalArticle https://hdl.handle.net/1721.1/133606 en 10.7554/ELIFE.60647 eLife Creative Commons Attribution 4.0 International license https://creativecommons.org/licenses/by/4.0/ application/pdf eLife Sciences Publications, Ltd eLife |
| spellingShingle | Shaaya, Mark Fauser, Jordan Zhurikhina, Anastasia Conage-Pough, Jason E Huyot, Vincent Brennan, Martin Flower, Cameron T Matsche, Jacob Khan, Shahzeb Natarajan, Viswanathan Rehman, Jalees Kota, Pradeep White, Forest M Tsygankov, Denis Karginov, Andrei V Light-regulated allosteric switch enables temporal and subcellular control of enzyme activity |
| title | Light-regulated allosteric switch enables temporal and subcellular control of enzyme activity |
| title_full | Light-regulated allosteric switch enables temporal and subcellular control of enzyme activity |
| title_fullStr | Light-regulated allosteric switch enables temporal and subcellular control of enzyme activity |
| title_full_unstemmed | Light-regulated allosteric switch enables temporal and subcellular control of enzyme activity |
| title_short | Light-regulated allosteric switch enables temporal and subcellular control of enzyme activity |
| title_sort | light regulated allosteric switch enables temporal and subcellular control of enzyme activity |
| url | https://hdl.handle.net/1721.1/133606 |
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