Delivery of the selenoprotein thioredoxin reductase 1 to mammalian cells
Over-expression of genetically encoded thioredoxin reductase 1 (TrxR1) TrxR1 can be toxic to cells due to the formation of a truncated version of the enzyme. We developed a new mammalian cell-based model to investigate TrxR1 activity. Fusion of the HIV-derived cell penetrating peptide (TAT) enabled...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2022-10-01
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| Series: | Frontiers in Molecular Biosciences |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/articles/10.3389/fmolb.2022.1031756/full |
| _version_ | 1811239581309206528 |
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| author | David E. Wright Tarana Siddika Ilka U. Heinemann Patrick O’Donoghue |
| author_facet | David E. Wright Tarana Siddika Ilka U. Heinemann Patrick O’Donoghue |
| author_sort | David E. Wright |
| collection | DOAJ |
| description | Over-expression of genetically encoded thioredoxin reductase 1 (TrxR1) TrxR1 can be toxic to cells due to the formation of a truncated version of the enzyme. We developed a new mammalian cell-based model to investigate TrxR1 activity. Fusion of the HIV-derived cell penetrating peptide (TAT) enabled efficient cellular uptake of purified TrxR1 containing 21 genetically encoded amino acids, including selenocysteine. The TAT peptide did not significantly alter the catalytic activity of TrxR1 in vitro. We monitored TrxR1-dependent redox activity in human cells using a TrxR1-specific red fluorescent live-cell reporter. Using programmed selenocysteine incorporation in Escherichia coli, our approach allowed efficient production of active recombinant human selenoprotein TrxR1 for delivery to the homologous context of the mammalian cell. The delivered TAT-TrxR1 showed robust activity in live cells and provided a novel platform to study TrxR1 biology in human cells. |
| first_indexed | 2024-04-12T13:03:32Z |
| format | Article |
| id | doaj.art-6123c34b77da47ce8c1831f4f11fcd5e |
| institution | Directory Open Access Journal |
| issn | 2296-889X |
| language | English |
| last_indexed | 2024-04-12T13:03:32Z |
| publishDate | 2022-10-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Molecular Biosciences |
| spelling | doaj.art-6123c34b77da47ce8c1831f4f11fcd5e2022-12-22T03:32:06ZengFrontiers Media S.A.Frontiers in Molecular Biosciences2296-889X2022-10-01910.3389/fmolb.2022.10317561031756Delivery of the selenoprotein thioredoxin reductase 1 to mammalian cellsDavid E. WrightTarana SiddikaIlka U. HeinemannPatrick O’DonoghueOver-expression of genetically encoded thioredoxin reductase 1 (TrxR1) TrxR1 can be toxic to cells due to the formation of a truncated version of the enzyme. We developed a new mammalian cell-based model to investigate TrxR1 activity. Fusion of the HIV-derived cell penetrating peptide (TAT) enabled efficient cellular uptake of purified TrxR1 containing 21 genetically encoded amino acids, including selenocysteine. The TAT peptide did not significantly alter the catalytic activity of TrxR1 in vitro. We monitored TrxR1-dependent redox activity in human cells using a TrxR1-specific red fluorescent live-cell reporter. Using programmed selenocysteine incorporation in Escherichia coli, our approach allowed efficient production of active recombinant human selenoprotein TrxR1 for delivery to the homologous context of the mammalian cell. The delivered TAT-TrxR1 showed robust activity in live cells and provided a novel platform to study TrxR1 biology in human cells.https://www.frontiersin.org/articles/10.3389/fmolb.2022.1031756/fullcell biologycell penetrating peptideenzymologygenetic code expansionredox biologyselenocysteine |
| spellingShingle | David E. Wright Tarana Siddika Ilka U. Heinemann Patrick O’Donoghue Delivery of the selenoprotein thioredoxin reductase 1 to mammalian cells Frontiers in Molecular Biosciences cell biology cell penetrating peptide enzymology genetic code expansion redox biology selenocysteine |
| title | Delivery of the selenoprotein thioredoxin reductase 1 to mammalian cells |
| title_full | Delivery of the selenoprotein thioredoxin reductase 1 to mammalian cells |
| title_fullStr | Delivery of the selenoprotein thioredoxin reductase 1 to mammalian cells |
| title_full_unstemmed | Delivery of the selenoprotein thioredoxin reductase 1 to mammalian cells |
| title_short | Delivery of the selenoprotein thioredoxin reductase 1 to mammalian cells |
| title_sort | delivery of the selenoprotein thioredoxin reductase 1 to mammalian cells |
| topic | cell biology cell penetrating peptide enzymology genetic code expansion redox biology selenocysteine |
| url | https://www.frontiersin.org/articles/10.3389/fmolb.2022.1031756/full |
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