Light-regulated allosteric switch enables temporal and subcellular control of enzyme activity
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 appro...
| Main Authors: | , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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eLife Sciences Publications Ltd
2020-09-01
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| Series: | eLife |
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| Online Access: | https://elifesciences.org/articles/60647 |
| _version_ | 1811236113018257408 |
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| author | Mark Shaaya Jordan Fauser Anastasia Zhurikhina Jason E Conage-Pough Vincent Huyot Martin Brennan Cameron T Flower Jacob Matsche Shahzeb Khan Viswanathan Natarajan Jalees Rehman Pradeep Kota Forest M White Denis Tsygankov Andrei V Karginov |
| author_facet | Mark Shaaya Jordan Fauser Anastasia Zhurikhina Jason E Conage-Pough Vincent Huyot Martin Brennan Cameron T Flower Jacob Matsche Shahzeb Khan Viswanathan Natarajan Jalees Rehman Pradeep Kota Forest M White Denis Tsygankov Andrei V Karginov |
| author_sort | Mark Shaaya |
| collection | DOAJ |
| description | 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 | 2024-04-12T12:03:20Z |
| format | Article |
| id | doaj.art-35f8a7f2365f48508aac921bafb81389 |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-12T12:03:20Z |
| publishDate | 2020-09-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-35f8a7f2365f48508aac921bafb813892022-12-22T03:33:46ZengeLife Sciences Publications LtdeLife2050-084X2020-09-01910.7554/eLife.60647Light-regulated allosteric switch enables temporal and subcellular control of enzyme activityMark Shaaya0https://orcid.org/0000-0003-4066-2693Jordan Fauser1https://orcid.org/0000-0002-0675-8977Anastasia Zhurikhina2Jason E Conage-Pough3https://orcid.org/0000-0002-1614-9374Vincent Huyot4Martin Brennan5Cameron T Flower6https://orcid.org/0000-0002-9632-9913Jacob Matsche7Shahzeb Khan8Viswanathan Natarajan9Jalees Rehman10https://orcid.org/0000-0002-2787-9292Pradeep Kota11Forest M White12https://orcid.org/0000-0002-1545-1651Denis Tsygankov13https://orcid.org/0000-0002-1180-3584Andrei V Karginov14https://orcid.org/0000-0003-2370-6383Department of Pharmacology and Regenerative Medicine, The University of Illinois at Chicago, College of Medicine, Chicago, United StatesDepartment of Pharmacology and Regenerative Medicine, The University of Illinois at Chicago, College of Medicine, Chicago, United StatesWallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University School of Medicine, Atlanta, United StatesThe David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, United States; Center for Precision Cancer Medicine, Massachusetts Institute of Technology, Cambridge, United States; Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, United StatesDepartment of Pharmacology and Regenerative Medicine, The University of Illinois at Chicago, College of Medicine, Chicago, United StatesDepartment of Pharmacology and Regenerative Medicine, The University of Illinois at Chicago, College of Medicine, Chicago, United StatesThe David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, United States; Center for Precision Cancer Medicine, Massachusetts Institute of Technology, Cambridge, United States; Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, United States; Program in Computational and Systems Biology, Massachusetts Institute of Technology, Cambridge, United StatesDepartment of Pharmacology and Regenerative Medicine, The University of Illinois at Chicago, College of Medicine, Chicago, United StatesDepartment of Pharmacology and Regenerative Medicine, The University of Illinois at Chicago, College of Medicine, Chicago, United StatesDepartment of Pharmacology and Regenerative Medicine, The University of Illinois at Chicago, College of Medicine, Chicago, United StatesDepartment of Pharmacology and Regenerative Medicine, The University of Illinois at Chicago, College of Medicine, Chicago, United States; University of Illinois Cancer Center, The University of Illinois at Chicago, Chicago, United States; Division of Cardiology, Department of Medicine, The University of Illinois, College of Medicine, Chicago, United StatesMarsico Lung Institute, Cystic Fibrosis Center and Department of Medicine, University of North Carolina, Chapel Hill, United StatesThe David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, United States; Center for Precision Cancer Medicine, Massachusetts Institute of Technology, Cambridge, United States; Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, United States; Program in Computational and Systems Biology, Massachusetts Institute of Technology, Cambridge, United StatesWallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University School of Medicine, Atlanta, United StatesDepartment of Pharmacology and Regenerative Medicine, The University of Illinois at Chicago, College of Medicine, Chicago, United States; University of Illinois Cancer Center, The University of Illinois at Chicago, Chicago, United StatesEngineered 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.https://elifesciences.org/articles/60647optogeneticsallostericenzymeprotein kinaseSrcprotein engineering |
| spellingShingle | Mark Shaaya Jordan Fauser Anastasia Zhurikhina Jason E Conage-Pough Vincent Huyot Martin Brennan Cameron T Flower Jacob Matsche Shahzeb Khan Viswanathan Natarajan Jalees Rehman Pradeep Kota Forest M White Denis Tsygankov Andrei V Karginov Light-regulated allosteric switch enables temporal and subcellular control of enzyme activity eLife optogenetics allosteric enzyme protein kinase Src protein engineering |
| 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 |
| topic | optogenetics allosteric enzyme protein kinase Src protein engineering |
| url | https://elifesciences.org/articles/60647 |
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