Nanobody-thioesterase chimeras to specifically target protein palmitoylation
Abstract The complexity of the cellular proteome is massively expanded by a repertoire of chemically distinct reversible post-translational modifications (PTMs) that control protein localisation, interactions, and function. The temporal and spatial control of these PTMs is central to organism physio...
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2025-02-01
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| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-025-56716-x |
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| author | Chien-Wen Kuo Caglar Gök Hannah Fulton Eleanor Dickson-Murray Samuel Adu Emily K. Gallen Sheon Mary Alan D. Robertson Fiona Jordan Emma Dunning William Mullen Godfrey L. Smith William Fuller |
| author_facet | Chien-Wen Kuo Caglar Gök Hannah Fulton Eleanor Dickson-Murray Samuel Adu Emily K. Gallen Sheon Mary Alan D. Robertson Fiona Jordan Emma Dunning William Mullen Godfrey L. Smith William Fuller |
| author_sort | Chien-Wen Kuo |
| collection | DOAJ |
| description | Abstract The complexity of the cellular proteome is massively expanded by a repertoire of chemically distinct reversible post-translational modifications (PTMs) that control protein localisation, interactions, and function. The temporal and spatial control of these PTMs is central to organism physiology, and mis-regulation of PTMs is a hallmark of many diseases. Here we present an approach to manipulate PTMs on target proteins using nanobodies fused to enzymes that control these PTMs. Anti-GFP nanobodies fused to thioesterases (which depalmitoylate protein cysteines) depalmitoylate GFP tagged substrates. A chemogenetic approach to enhance nanobody affinity for its target enables temporal control of target depalmitoylation. Using a thioesterase fused to a nanobody directed against the Ca(v)1.2 beta subunit we reduce palmitoylation of the Ca(v)1.2 alpha subunit, modifying the channel’s voltage dependence and arrhythmia susceptibility in stem cell derived cardiac myocytes. We conclude that nanobody enzyme chimeras represent an approach to specifically manipulate PTMs, with applications in both the laboratory and the clinic. |
| format | Article |
| id | doaj-art-4de661ede5f34506bff5da0942e57cdb |
| institution | Kabale University |
| issn | 2041-1723 |
| language | English |
| publishDate | 2025-02-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj-art-4de661ede5f34506bff5da0942e57cdb2025-02-09T12:45:39ZengNature PortfolioNature Communications2041-17232025-02-0116111310.1038/s41467-025-56716-xNanobody-thioesterase chimeras to specifically target protein palmitoylationChien-Wen Kuo0Caglar Gök1Hannah Fulton2Eleanor Dickson-Murray3Samuel Adu4Emily K. Gallen5Sheon Mary6Alan D. Robertson7Fiona Jordan8Emma Dunning9William Mullen10Godfrey L. Smith11William Fuller12School of Cardiovascular & Metabolic Health, College of Medical Veterinary and Life Sciences, University of GlasgowSchool of Cardiovascular & Metabolic Health, College of Medical Veterinary and Life Sciences, University of GlasgowSchool of Cardiovascular & Metabolic Health, College of Medical Veterinary and Life Sciences, University of GlasgowSchool of Cardiovascular & Metabolic Health, College of Medical Veterinary and Life Sciences, University of GlasgowSchool of Cardiovascular & Metabolic Health, College of Medical Veterinary and Life Sciences, University of GlasgowSchool of Cardiovascular & Metabolic Health, College of Medical Veterinary and Life Sciences, University of GlasgowSchool of Cardiovascular & Metabolic Health, College of Medical Veterinary and Life Sciences, University of GlasgowSchool of Cardiovascular & Metabolic Health, College of Medical Veterinary and Life Sciences, University of GlasgowSchool of Cardiovascular & Metabolic Health, College of Medical Veterinary and Life Sciences, University of GlasgowSchool of Cardiovascular & Metabolic Health, College of Medical Veterinary and Life Sciences, University of GlasgowSchool of Cardiovascular & Metabolic Health, College of Medical Veterinary and Life Sciences, University of GlasgowSchool of Cardiovascular & Metabolic Health, College of Medical Veterinary and Life Sciences, University of GlasgowSchool of Cardiovascular & Metabolic Health, College of Medical Veterinary and Life Sciences, University of GlasgowAbstract The complexity of the cellular proteome is massively expanded by a repertoire of chemically distinct reversible post-translational modifications (PTMs) that control protein localisation, interactions, and function. The temporal and spatial control of these PTMs is central to organism physiology, and mis-regulation of PTMs is a hallmark of many diseases. Here we present an approach to manipulate PTMs on target proteins using nanobodies fused to enzymes that control these PTMs. Anti-GFP nanobodies fused to thioesterases (which depalmitoylate protein cysteines) depalmitoylate GFP tagged substrates. A chemogenetic approach to enhance nanobody affinity for its target enables temporal control of target depalmitoylation. Using a thioesterase fused to a nanobody directed against the Ca(v)1.2 beta subunit we reduce palmitoylation of the Ca(v)1.2 alpha subunit, modifying the channel’s voltage dependence and arrhythmia susceptibility in stem cell derived cardiac myocytes. We conclude that nanobody enzyme chimeras represent an approach to specifically manipulate PTMs, with applications in both the laboratory and the clinic.https://doi.org/10.1038/s41467-025-56716-x |
| spellingShingle | Chien-Wen Kuo Caglar Gök Hannah Fulton Eleanor Dickson-Murray Samuel Adu Emily K. Gallen Sheon Mary Alan D. Robertson Fiona Jordan Emma Dunning William Mullen Godfrey L. Smith William Fuller Nanobody-thioesterase chimeras to specifically target protein palmitoylation Nature Communications |
| title | Nanobody-thioesterase chimeras to specifically target protein palmitoylation |
| title_full | Nanobody-thioesterase chimeras to specifically target protein palmitoylation |
| title_fullStr | Nanobody-thioesterase chimeras to specifically target protein palmitoylation |
| title_full_unstemmed | Nanobody-thioesterase chimeras to specifically target protein palmitoylation |
| title_short | Nanobody-thioesterase chimeras to specifically target protein palmitoylation |
| title_sort | nanobody thioesterase chimeras to specifically target protein palmitoylation |
| url | https://doi.org/10.1038/s41467-025-56716-x |
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