The Extracellular Domain of Human High Affinity Copper Transporter (hNdCTR1), Synthesized by E. coli Cells, Chelates Silver and Copper Ions In Vivo
There is much interest in effective copper chelators to correct copper dyshomeostasis in neurodegenerative and oncological diseases. In this study, a recombinant fusion protein for expression in Escherichia coli cells was constructed from glutathione-S-transferase (GST) and the N-terminal domain (ec...
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| Format: | Article |
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MDPI AG
2017-11-01
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| Series: | Biomolecules |
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| Online Access: | https://www.mdpi.com/2218-273X/7/4/78 |
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| author | Tatiana P. Sankova Iurii A. Orlov Andrey N. Saveliev Demid A. Kirilenko Polina S. Babich Pavel N. Brunkov Ludmila V. Puchkova |
| author_facet | Tatiana P. Sankova Iurii A. Orlov Andrey N. Saveliev Demid A. Kirilenko Polina S. Babich Pavel N. Brunkov Ludmila V. Puchkova |
| author_sort | Tatiana P. Sankova |
| collection | DOAJ |
| description | There is much interest in effective copper chelators to correct copper dyshomeostasis in neurodegenerative and oncological diseases. In this study, a recombinant fusion protein for expression in Escherichia coli cells was constructed from glutathione-S-transferase (GST) and the N-terminal domain (ectodomain) of human high affinity copper transporter CTR1 (hNdCTR1), which has three metal-bound motifs. Several biological properties of the GST-hNdCTR1 fusion protein were assessed. It was demonstrated that in cells, the protein was prone to oligomerization, formed inclusion bodies and displayed no toxicity. Treatment of E. coli cells with copper and silver ions reduced cell viability in a dose- and time-dependent manner. Cells expressing GST-hNdCTR1 protein demonstrated resistance to the metal treatments. These cells accumulated silver ions and formed nanoparticles that contained AgCl and metallic silver. In this bacterial population, filamentous bacteria with a length of about 10 µm were often observed. The possibility for the fusion protein carrying extracellular metal binding motifs to integrate into the cell’s copper metabolism and its chelating properties are discussed. |
| first_indexed | 2024-12-22T15:28:09Z |
| format | Article |
| id | doaj.art-ab79763d996f465b87f4bc35c9cb92ed |
| institution | Directory Open Access Journal |
| issn | 2218-273X |
| language | English |
| last_indexed | 2024-12-22T15:28:09Z |
| publishDate | 2017-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biomolecules |
| spelling | doaj.art-ab79763d996f465b87f4bc35c9cb92ed2022-12-21T18:21:26ZengMDPI AGBiomolecules2218-273X2017-11-01747810.3390/biom7040078biom7040078The Extracellular Domain of Human High Affinity Copper Transporter (hNdCTR1), Synthesized by E. coli Cells, Chelates Silver and Copper Ions In VivoTatiana P. Sankova0Iurii A. Orlov1Andrey N. Saveliev2Demid A. Kirilenko3Polina S. Babich4Pavel N. Brunkov5Ludmila V. Puchkova6Department of Biophysics, Peter the Great St. Petersburg Polytechnic University, Politekhnicheskaya str., 29, St.-Petersburg 195251, RussiaDepartment of Biophysics, Peter the Great St. Petersburg Polytechnic University, Politekhnicheskaya str., 29, St.-Petersburg 195251, RussiaDepartment of Biophysics, Peter the Great St. Petersburg Polytechnic University, Politekhnicheskaya str., 29, St.-Petersburg 195251, RussiaDepartment of Modern Functional Materials, ITMO University, Kronverksky av., 49, St.-Petersburg 197101, RussiaDepartment of Zoology, Herzen State Pedagogical University of Russia, Kazanskaya str., 6, St.-Petersburg 191186, RussiaDepartment of Modern Functional Materials, ITMO University, Kronverksky av., 49, St.-Petersburg 197101, RussiaDepartment of Biophysics, Peter the Great St. Petersburg Polytechnic University, Politekhnicheskaya str., 29, St.-Petersburg 195251, RussiaThere is much interest in effective copper chelators to correct copper dyshomeostasis in neurodegenerative and oncological diseases. In this study, a recombinant fusion protein for expression in Escherichia coli cells was constructed from glutathione-S-transferase (GST) and the N-terminal domain (ectodomain) of human high affinity copper transporter CTR1 (hNdCTR1), which has three metal-bound motifs. Several biological properties of the GST-hNdCTR1 fusion protein were assessed. It was demonstrated that in cells, the protein was prone to oligomerization, formed inclusion bodies and displayed no toxicity. Treatment of E. coli cells with copper and silver ions reduced cell viability in a dose- and time-dependent manner. Cells expressing GST-hNdCTR1 protein demonstrated resistance to the metal treatments. These cells accumulated silver ions and formed nanoparticles that contained AgCl and metallic silver. In this bacterial population, filamentous bacteria with a length of about 10 µm were often observed. The possibility for the fusion protein carrying extracellular metal binding motifs to integrate into the cell’s copper metabolism and its chelating properties are discussed.https://www.mdpi.com/2218-273X/7/4/78copper transporter 1 metal-binding extracellular domain cloningcopper/silver chelationEscherichia coli filamentous growthsecondary silver nanoparticles formation |
| spellingShingle | Tatiana P. Sankova Iurii A. Orlov Andrey N. Saveliev Demid A. Kirilenko Polina S. Babich Pavel N. Brunkov Ludmila V. Puchkova The Extracellular Domain of Human High Affinity Copper Transporter (hNdCTR1), Synthesized by E. coli Cells, Chelates Silver and Copper Ions In Vivo Biomolecules copper transporter 1 metal-binding extracellular domain cloning copper/silver chelation Escherichia coli filamentous growth secondary silver nanoparticles formation |
| title | The Extracellular Domain of Human High Affinity Copper Transporter (hNdCTR1), Synthesized by E. coli Cells, Chelates Silver and Copper Ions In Vivo |
| title_full | The Extracellular Domain of Human High Affinity Copper Transporter (hNdCTR1), Synthesized by E. coli Cells, Chelates Silver and Copper Ions In Vivo |
| title_fullStr | The Extracellular Domain of Human High Affinity Copper Transporter (hNdCTR1), Synthesized by E. coli Cells, Chelates Silver and Copper Ions In Vivo |
| title_full_unstemmed | The Extracellular Domain of Human High Affinity Copper Transporter (hNdCTR1), Synthesized by E. coli Cells, Chelates Silver and Copper Ions In Vivo |
| title_short | The Extracellular Domain of Human High Affinity Copper Transporter (hNdCTR1), Synthesized by E. coli Cells, Chelates Silver and Copper Ions In Vivo |
| title_sort | extracellular domain of human high affinity copper transporter hndctr1 synthesized by e coli cells chelates silver and copper ions in vivo |
| topic | copper transporter 1 metal-binding extracellular domain cloning copper/silver chelation Escherichia coli filamentous growth secondary silver nanoparticles formation |
| url | https://www.mdpi.com/2218-273X/7/4/78 |
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