Engineering a switchable single‐chain TEV protease to control protein maturation in living neurons
Abstract Engineered proteases are promising tools to address physiological and pathophysiological questions as well as to develop new therapeutic approaches. Here we introduce a new genetically encoded engineered single‐chain tobacco etch virus protease, allowing to control proprotein cleavage in di...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Wiley
2022-05-01
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| Series: | Bioengineering & Translational Medicine |
| Subjects: | |
| Online Access: | https://doi.org/10.1002/btm2.10292 |
| _version_ | 1828553750283812864 |
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| author | Pietro Renna Cristian Ripoli Onur Dagliyan Francesco Pastore Marco Rinaudo Agnese Re Fabiola Paciello Claudio Grassi |
| author_facet | Pietro Renna Cristian Ripoli Onur Dagliyan Francesco Pastore Marco Rinaudo Agnese Re Fabiola Paciello Claudio Grassi |
| author_sort | Pietro Renna |
| collection | DOAJ |
| description | Abstract Engineered proteases are promising tools to address physiological and pathophysiological questions as well as to develop new therapeutic approaches. Here we introduce a new genetically encoded engineered single‐chain tobacco etch virus protease, allowing to control proprotein cleavage in different compartments of living mammalian cells. We demonstrated a set of controllable proteolytic effects, including cytosolic protein cleavage, inducible gene expression, and maturation of brain‐derived neurotrophic factor (BDNF) in the secretory pathway thus showing the versatility of this technique. Of note, the secretory pathway exhibits different characteristics from the cytosol and it is difficult to target because inaccessible to some small molecules. We were able to induce ligand‐mediated BDNF maturation and monitor its effects on dendritic spines in hippocampal pyramidal cells and in the mouse brain. This strategy paves the way to dissect proteolytic cleavage product signaling in various processes as well as for future therapeutic applications. |
| first_indexed | 2024-12-12T05:24:04Z |
| format | Article |
| id | doaj.art-36c7ecebe40c4f0286e53ec17c30509c |
| institution | Directory Open Access Journal |
| issn | 2380-6761 |
| language | English |
| last_indexed | 2024-12-12T05:24:04Z |
| publishDate | 2022-05-01 |
| publisher | Wiley |
| record_format | Article |
| series | Bioengineering & Translational Medicine |
| spelling | doaj.art-36c7ecebe40c4f0286e53ec17c30509c2022-12-22T00:36:32ZengWileyBioengineering & Translational Medicine2380-67612022-05-0172n/an/a10.1002/btm2.10292Engineering a switchable single‐chain TEV protease to control protein maturation in living neuronsPietro Renna0Cristian Ripoli1Onur Dagliyan2Francesco Pastore3Marco Rinaudo4Agnese Re5Fabiola Paciello6Claudio Grassi7Department of Neuroscience Università Cattolica del Sacro Cuore Rome ItalyDepartment of Neuroscience Università Cattolica del Sacro Cuore Rome ItalyDepartment of Neurobiology Harvard Medical School Boston Massachusetts USADepartment of Neuroscience Università Cattolica del Sacro Cuore Rome ItalyDepartment of Neuroscience Università Cattolica del Sacro Cuore Rome ItalyDepartment of Neuroscience Università Cattolica del Sacro Cuore Rome ItalyDepartment of Neuroscience Università Cattolica del Sacro Cuore Rome ItalyDepartment of Neuroscience Università Cattolica del Sacro Cuore Rome ItalyAbstract Engineered proteases are promising tools to address physiological and pathophysiological questions as well as to develop new therapeutic approaches. Here we introduce a new genetically encoded engineered single‐chain tobacco etch virus protease, allowing to control proprotein cleavage in different compartments of living mammalian cells. We demonstrated a set of controllable proteolytic effects, including cytosolic protein cleavage, inducible gene expression, and maturation of brain‐derived neurotrophic factor (BDNF) in the secretory pathway thus showing the versatility of this technique. Of note, the secretory pathway exhibits different characteristics from the cytosol and it is difficult to target because inaccessible to some small molecules. We were able to induce ligand‐mediated BDNF maturation and monitor its effects on dendritic spines in hippocampal pyramidal cells and in the mouse brain. This strategy paves the way to dissect proteolytic cleavage product signaling in various processes as well as for future therapeutic applications.https://doi.org/10.1002/btm2.10292dendritic spinesneuronal plasticityneurotrophinsproteaseprotein engineeringTEV |
| spellingShingle | Pietro Renna Cristian Ripoli Onur Dagliyan Francesco Pastore Marco Rinaudo Agnese Re Fabiola Paciello Claudio Grassi Engineering a switchable single‐chain TEV protease to control protein maturation in living neurons Bioengineering & Translational Medicine dendritic spines neuronal plasticity neurotrophins protease protein engineering TEV |
| title | Engineering a switchable single‐chain TEV protease to control protein maturation in living neurons |
| title_full | Engineering a switchable single‐chain TEV protease to control protein maturation in living neurons |
| title_fullStr | Engineering a switchable single‐chain TEV protease to control protein maturation in living neurons |
| title_full_unstemmed | Engineering a switchable single‐chain TEV protease to control protein maturation in living neurons |
| title_short | Engineering a switchable single‐chain TEV protease to control protein maturation in living neurons |
| title_sort | engineering a switchable single chain tev protease to control protein maturation in living neurons |
| topic | dendritic spines neuronal plasticity neurotrophins protease protein engineering TEV |
| url | https://doi.org/10.1002/btm2.10292 |
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