Aptamer-Based Imaging of Polyisoprenoids in the Malaria Parasite
Dolichols are isoprenoid end-products of the mevalonate and 2<i>C</i>-methyl-D-erythritol-4-phosphate pathways. The synthesis of dolichols is initiated with the addition of several molecules of isopentenyl diphosphate to farnesyl diphosphate. This reaction is catalyzed by a <i>cis&...
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MDPI AG
2023-12-01
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| author | Flavia M. Zimbres Emilio F. Merino Grant J. Butschek Joshua H. Butler Frédéric Ducongé Maria B. Cassera |
| author_facet | Flavia M. Zimbres Emilio F. Merino Grant J. Butschek Joshua H. Butler Frédéric Ducongé Maria B. Cassera |
| author_sort | Flavia M. Zimbres |
| collection | DOAJ |
| description | Dolichols are isoprenoid end-products of the mevalonate and 2<i>C</i>-methyl-D-erythritol-4-phosphate pathways. The synthesis of dolichols is initiated with the addition of several molecules of isopentenyl diphosphate to farnesyl diphosphate. This reaction is catalyzed by a <i>cis</i>-prenyltransferase and leads to the formation of polyprenyl diphosphate. Subsequent steps involve the dephosphorylation and reduction of the α-isoprene unit by a polyprenol reductase, resulting in the generation of dolichol. The size of the dolichol varies, depending on the number of isoprene units incorporated. In eukaryotes, dolichols are synthesized as a mixture of four or more different lengths. Their biosynthesis is predicted to occur in the endoplasmic reticulum, where dolichols play an essential role in protein glycosylation. In this study, we have developed a selection of aptamers targeting dolichols and enhanced their specificity by incorporating fatty acids for negative selection. One aptamer showed high enrichment and specificity for linear polyisoprenoids containing at least one oxygen atom, such as an alcohol or aldehyde, in the α-isoprene unit. The selected aptamer proved to be a valuable tool for the subcellular localization of polyisoprenoids in the malaria parasite. To the best of our knowledge, this is the first time that polyisoprenoids have been localized within a cell using aptamer-based imaging techniques. |
| first_indexed | 2024-03-08T15:01:01Z |
| format | Article |
| id | doaj.art-0762b8beb3464a489c44762777b2e273 |
| institution | Directory Open Access Journal |
| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-03-08T15:01:01Z |
| publishDate | 2023-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Molecules |
| spelling | doaj.art-0762b8beb3464a489c44762777b2e2732024-01-10T15:04:24ZengMDPI AGMolecules1420-30492023-12-0129117810.3390/molecules29010178Aptamer-Based Imaging of Polyisoprenoids in the Malaria ParasiteFlavia M. Zimbres0Emilio F. Merino1Grant J. Butschek2Joshua H. Butler3Frédéric Ducongé4Maria B. Cassera5Department of Biochemistry and Molecular Biology, University of Georgia, Athens, GA 30602, USADepartment of Biochemistry and Molecular Biology, University of Georgia, Athens, GA 30602, USADepartment of Biochemistry and Molecular Biology, University of Georgia, Athens, GA 30602, USADepartment of Biochemistry and Molecular Biology, University of Georgia, Athens, GA 30602, USAFrench Atomic Energy Commission (CEA), Fundamental Research Division (DRF), Institute of Biology François Jacob (Jacob), Molecular Imaging Research Center, 92265 Fontenay-aux-Roses, FranceDepartment of Biochemistry and Molecular Biology, University of Georgia, Athens, GA 30602, USADolichols are isoprenoid end-products of the mevalonate and 2<i>C</i>-methyl-D-erythritol-4-phosphate pathways. The synthesis of dolichols is initiated with the addition of several molecules of isopentenyl diphosphate to farnesyl diphosphate. This reaction is catalyzed by a <i>cis</i>-prenyltransferase and leads to the formation of polyprenyl diphosphate. Subsequent steps involve the dephosphorylation and reduction of the α-isoprene unit by a polyprenol reductase, resulting in the generation of dolichol. The size of the dolichol varies, depending on the number of isoprene units incorporated. In eukaryotes, dolichols are synthesized as a mixture of four or more different lengths. Their biosynthesis is predicted to occur in the endoplasmic reticulum, where dolichols play an essential role in protein glycosylation. In this study, we have developed a selection of aptamers targeting dolichols and enhanced their specificity by incorporating fatty acids for negative selection. One aptamer showed high enrichment and specificity for linear polyisoprenoids containing at least one oxygen atom, such as an alcohol or aldehyde, in the α-isoprene unit. The selected aptamer proved to be a valuable tool for the subcellular localization of polyisoprenoids in the malaria parasite. To the best of our knowledge, this is the first time that polyisoprenoids have been localized within a cell using aptamer-based imaging techniques.https://www.mdpi.com/1420-3049/29/1/178<i>Plasmodium</i>polyisoprenoidsaptamersaptamer-based imaging |
| spellingShingle | Flavia M. Zimbres Emilio F. Merino Grant J. Butschek Joshua H. Butler Frédéric Ducongé Maria B. Cassera Aptamer-Based Imaging of Polyisoprenoids in the Malaria Parasite Molecules <i>Plasmodium</i> polyisoprenoids aptamers aptamer-based imaging |
| title | Aptamer-Based Imaging of Polyisoprenoids in the Malaria Parasite |
| title_full | Aptamer-Based Imaging of Polyisoprenoids in the Malaria Parasite |
| title_fullStr | Aptamer-Based Imaging of Polyisoprenoids in the Malaria Parasite |
| title_full_unstemmed | Aptamer-Based Imaging of Polyisoprenoids in the Malaria Parasite |
| title_short | Aptamer-Based Imaging of Polyisoprenoids in the Malaria Parasite |
| title_sort | aptamer based imaging of polyisoprenoids in the malaria parasite |
| topic | <i>Plasmodium</i> polyisoprenoids aptamers aptamer-based imaging |
| url | https://www.mdpi.com/1420-3049/29/1/178 |
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