A metal-trap tests and refines blueprints to engineer cellular protein metalation with different elements
Abstract It has been challenging to test how proteins acquire specific metals in cells. The speciation of metalation is thought to depend on the preferences of proteins for different metals competing at intracellular metal-availabilities. This implies mis-metalation may occur if proteins become mis-...
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2025-01-01
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| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-025-56199-w |
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| author | Sophie E. Clough Tessa R. Young Emma Tarrant Andrew J. P. Scott Peter T. Chivers Arthur Glasfeld Nigel J. Robinson |
| author_facet | Sophie E. Clough Tessa R. Young Emma Tarrant Andrew J. P. Scott Peter T. Chivers Arthur Glasfeld Nigel J. Robinson |
| author_sort | Sophie E. Clough |
| collection | DOAJ |
| description | Abstract It has been challenging to test how proteins acquire specific metals in cells. The speciation of metalation is thought to depend on the preferences of proteins for different metals competing at intracellular metal-availabilities. This implies mis-metalation may occur if proteins become mis-matched to metal-availabilities in heterologous cells. Here we use a cyanobacterial MnII-cupin (MncA) as a metal trap, to test predictions of metalation. By re-folding MncA in buffered competing metals, metal-preferences are determined. Relating metal-preferences to metal-availabilities estimated using cellular metal sensors, predicts mis-metalation of MncA with FeII in E. coli. After expression in E. coli, predominantly FeII-bound MncA is isolated experimentally. It is predicted that in metal-supplemented viable cells metal-MncA speciation should switch. MnII-, CoII-, or NiII-MncA are recovered from the respective metal-supplemented cells. Differences between observed and predicted metal-MncA speciation are used to refine estimated metal availabilities. Values are provided as blueprints to guide engineering biological protein metalation. |
| format | Article |
| id | doaj-art-b5e6def0f9d4465ba16da85d665a013f |
| institution | Kabale University |
| issn | 2041-1723 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Nature Portfolio |
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| series | Nature Communications |
| spelling | doaj-art-b5e6def0f9d4465ba16da85d665a013f2025-01-19T12:32:13ZengNature PortfolioNature Communications2041-17232025-01-0116111510.1038/s41467-025-56199-wA metal-trap tests and refines blueprints to engineer cellular protein metalation with different elementsSophie E. Clough0Tessa R. Young1Emma Tarrant2Andrew J. P. Scott3Peter T. Chivers4Arthur Glasfeld5Nigel J. Robinson6Department of Biosciences, University of DurhamDepartment of Biosciences, University of DurhamDepartment of Biosciences, University of DurhamDepartment of Biosciences, University of DurhamDepartment of Biosciences, University of DurhamDepartment of Biosciences, University of DurhamDepartment of Biosciences, University of DurhamAbstract It has been challenging to test how proteins acquire specific metals in cells. The speciation of metalation is thought to depend on the preferences of proteins for different metals competing at intracellular metal-availabilities. This implies mis-metalation may occur if proteins become mis-matched to metal-availabilities in heterologous cells. Here we use a cyanobacterial MnII-cupin (MncA) as a metal trap, to test predictions of metalation. By re-folding MncA in buffered competing metals, metal-preferences are determined. Relating metal-preferences to metal-availabilities estimated using cellular metal sensors, predicts mis-metalation of MncA with FeII in E. coli. After expression in E. coli, predominantly FeII-bound MncA is isolated experimentally. It is predicted that in metal-supplemented viable cells metal-MncA speciation should switch. MnII-, CoII-, or NiII-MncA are recovered from the respective metal-supplemented cells. Differences between observed and predicted metal-MncA speciation are used to refine estimated metal availabilities. Values are provided as blueprints to guide engineering biological protein metalation.https://doi.org/10.1038/s41467-025-56199-w |
| spellingShingle | Sophie E. Clough Tessa R. Young Emma Tarrant Andrew J. P. Scott Peter T. Chivers Arthur Glasfeld Nigel J. Robinson A metal-trap tests and refines blueprints to engineer cellular protein metalation with different elements Nature Communications |
| title | A metal-trap tests and refines blueprints to engineer cellular protein metalation with different elements |
| title_full | A metal-trap tests and refines blueprints to engineer cellular protein metalation with different elements |
| title_fullStr | A metal-trap tests and refines blueprints to engineer cellular protein metalation with different elements |
| title_full_unstemmed | A metal-trap tests and refines blueprints to engineer cellular protein metalation with different elements |
| title_short | A metal-trap tests and refines blueprints to engineer cellular protein metalation with different elements |
| title_sort | metal trap tests and refines blueprints to engineer cellular protein metalation with different elements |
| url | https://doi.org/10.1038/s41467-025-56199-w |
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