Construction of an artificial cell membrane anchor using DARC as a fitting for artificial extracellular functionalities of eukaryotic cells
<p>Abstract</p> <p>The need to functionalize cell membranes in a directed way for specific applications as single cell arrays or to force close cell-to-cell contact for artificial intercellular interaction and/or induction concerning stem cell manipulation or in general to have a t...
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| Format: | Article |
| Language: | English |
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BMC
2012-01-01
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| Series: | Journal of Nanobiotechnology |
| Subjects: | |
| Online Access: | http://www.jnanobiotechnology.com/content/10/1/1 |
| _version_ | 1828282546232754176 |
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| author | von Nickisch-Rosenegk Markus Teschke Till Bier Frank F |
| author_facet | von Nickisch-Rosenegk Markus Teschke Till Bier Frank F |
| author_sort | von Nickisch-Rosenegk Markus |
| collection | DOAJ |
| description | <p>Abstract</p> <p>The need to functionalize cell membranes in a directed way for specific applications as single cell arrays or to force close cell-to-cell contact for artificial intercellular interaction and/or induction concerning stem cell manipulation or in general to have a tool for membrane and cell surface-associated processes, we envisaged a neutral inactive membrane anchor for extracellular entities to facillitate the above mentioned functionalities.</p> <p>The silent Duffy antigen/receptor for chemokines (DARC) is a receptor-like membrane protein of erythrocytes and mediates no cell transduction not at least regarding a missing or truncated G-loop and therefore it seemed to be the candidate for our cell membrane anchor.</p> <p>We isolated the genetic information of DARC from human genomic DNA and cloned it in a mammalian cell line as a fusion protein via a suitable plasmid vector.</p> <p>In this report we demonstrate that the human plasma membrane protein DARC can be used as an artificial anchor molecule in cell surface engineering applications. We constructed the fusion protein SNAP-tag-DARC, consisting of DARC and the self-labeling protein tag SNAP-tag<sup>® </sup>(Covalys). The SNAP-tag® served as an example for a molecular-technological developed protein that is artificially attached to the extracellular side of the plasma membrane through our DARC-anchor. SnapTag should serve as an example for any extracellular entity and was easy to detect by a commercial detection system. The synthesis of SNAP-tag-DARC, its correct incorporation into the cell membrane and the functionality of the SNAP-tag® were verified by RT-PCR, Western blotting and confocal fluorescence microscopy and showed the desired functionality as an membrane anchor for an extracellular application entity.</p> |
| first_indexed | 2024-04-13T08:31:14Z |
| format | Article |
| id | doaj.art-7c64fbe36f6e47319da360dc56c72b89 |
| institution | Directory Open Access Journal |
| issn | 1477-3155 |
| language | English |
| last_indexed | 2024-04-13T08:31:14Z |
| publishDate | 2012-01-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Nanobiotechnology |
| spelling | doaj.art-7c64fbe36f6e47319da360dc56c72b892022-12-22T02:54:15ZengBMCJournal of Nanobiotechnology1477-31552012-01-01101110.1186/1477-3155-10-1Construction of an artificial cell membrane anchor using DARC as a fitting for artificial extracellular functionalities of eukaryotic cellsvon Nickisch-Rosenegk MarkusTeschke TillBier Frank F<p>Abstract</p> <p>The need to functionalize cell membranes in a directed way for specific applications as single cell arrays or to force close cell-to-cell contact for artificial intercellular interaction and/or induction concerning stem cell manipulation or in general to have a tool for membrane and cell surface-associated processes, we envisaged a neutral inactive membrane anchor for extracellular entities to facillitate the above mentioned functionalities.</p> <p>The silent Duffy antigen/receptor for chemokines (DARC) is a receptor-like membrane protein of erythrocytes and mediates no cell transduction not at least regarding a missing or truncated G-loop and therefore it seemed to be the candidate for our cell membrane anchor.</p> <p>We isolated the genetic information of DARC from human genomic DNA and cloned it in a mammalian cell line as a fusion protein via a suitable plasmid vector.</p> <p>In this report we demonstrate that the human plasma membrane protein DARC can be used as an artificial anchor molecule in cell surface engineering applications. We constructed the fusion protein SNAP-tag-DARC, consisting of DARC and the self-labeling protein tag SNAP-tag<sup>® </sup>(Covalys). The SNAP-tag® served as an example for a molecular-technological developed protein that is artificially attached to the extracellular side of the plasma membrane through our DARC-anchor. SnapTag should serve as an example for any extracellular entity and was easy to detect by a commercial detection system. The synthesis of SNAP-tag-DARC, its correct incorporation into the cell membrane and the functionality of the SNAP-tag® were verified by RT-PCR, Western blotting and confocal fluorescence microscopy and showed the desired functionality as an membrane anchor for an extracellular application entity.</p>http://www.jnanobiotechnology.com/content/10/1/1DuffyDARCartificial cell membrane anchorartificial cell membrane functionalitiesrecombinant functional membrane fusion protein |
| spellingShingle | von Nickisch-Rosenegk Markus Teschke Till Bier Frank F Construction of an artificial cell membrane anchor using DARC as a fitting for artificial extracellular functionalities of eukaryotic cells Journal of Nanobiotechnology Duffy DARC artificial cell membrane anchor artificial cell membrane functionalities recombinant functional membrane fusion protein |
| title | Construction of an artificial cell membrane anchor using DARC as a fitting for artificial extracellular functionalities of eukaryotic cells |
| title_full | Construction of an artificial cell membrane anchor using DARC as a fitting for artificial extracellular functionalities of eukaryotic cells |
| title_fullStr | Construction of an artificial cell membrane anchor using DARC as a fitting for artificial extracellular functionalities of eukaryotic cells |
| title_full_unstemmed | Construction of an artificial cell membrane anchor using DARC as a fitting for artificial extracellular functionalities of eukaryotic cells |
| title_short | Construction of an artificial cell membrane anchor using DARC as a fitting for artificial extracellular functionalities of eukaryotic cells |
| title_sort | construction of an artificial cell membrane anchor using darc as a fitting for artificial extracellular functionalities of eukaryotic cells |
| topic | Duffy DARC artificial cell membrane anchor artificial cell membrane functionalities recombinant functional membrane fusion protein |
| url | http://www.jnanobiotechnology.com/content/10/1/1 |
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