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Selection of an Aptamer against the Enzyme 1-deoxy-D-xylulose-5-phosphate Reductoisomerase from Plasmodium falciparum

The methyl erythritol phosphate (MEP) pathway of isoprenoid biosynthesis is essential for malaria parasites and also for several human pathogenic bacteria, thus representing an interesting target for future antimalarials and antibiotics and for diagnostic strategies. We have developed a DNA aptamer...

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Autores principales: Roca, Carlota, Avalos-Padilla, Yunuen, Prieto-Simón, Beatriz, Iglesias, Valentín, Ramírez, Miriam, Imperial, Santiago, Fernàndez-Busquets, Xavier
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9695703/
https://www.ncbi.nlm.nih.gov/pubmed/36432706
http://dx.doi.org/10.3390/pharmaceutics14112515
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author Roca, Carlota
Avalos-Padilla, Yunuen
Prieto-Simón, Beatriz
Iglesias, Valentín
Ramírez, Miriam
Imperial, Santiago
Fernàndez-Busquets, Xavier
author_facet Roca, Carlota
Avalos-Padilla, Yunuen
Prieto-Simón, Beatriz
Iglesias, Valentín
Ramírez, Miriam
Imperial, Santiago
Fernàndez-Busquets, Xavier
author_sort Roca, Carlota
collection PubMed
description The methyl erythritol phosphate (MEP) pathway of isoprenoid biosynthesis is essential for malaria parasites and also for several human pathogenic bacteria, thus representing an interesting target for future antimalarials and antibiotics and for diagnostic strategies. We have developed a DNA aptamer (D10) against Plasmodium falciparum 1-deoxy-D-xylulose-5-phosphate reductoisomerase (DXR), the second enzyme of this metabolic route. D10 binds in vitro to recombinant DXR from P. falciparum and Escherichia coli, showing at 10 µM a ca. 50% inhibition of the bacterial enzyme. In silico docking analysis indicates that D10 associates with DXR in solvent-exposed regions outside the active center pocket. According to fluorescence confocal microscopy data, this aptamer specifically targets in P. falciparum in vitro cultures the apicoplast organelle where the MEP pathway is localized and is, therefore, a highly specific marker of red blood cells parasitized by Plasmodium vs. naïve erythrocytes. D10 is also selective for the detection of MEP+ bacteria (e.g., E. coli and Pseudomonas aeruginosa) vs. those lacking DXR (e.g., Enterococcus faecalis). Based on these results, we discuss the potential of DNA aptamers in the development of ligands that can outcompete the performance of the well-established antibody technology for future therapeutic and diagnostic approaches.
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spelling pubmed-96957032022-11-26 Selection of an Aptamer against the Enzyme 1-deoxy-D-xylulose-5-phosphate Reductoisomerase from Plasmodium falciparum Roca, Carlota Avalos-Padilla, Yunuen Prieto-Simón, Beatriz Iglesias, Valentín Ramírez, Miriam Imperial, Santiago Fernàndez-Busquets, Xavier Pharmaceutics Article The methyl erythritol phosphate (MEP) pathway of isoprenoid biosynthesis is essential for malaria parasites and also for several human pathogenic bacteria, thus representing an interesting target for future antimalarials and antibiotics and for diagnostic strategies. We have developed a DNA aptamer (D10) against Plasmodium falciparum 1-deoxy-D-xylulose-5-phosphate reductoisomerase (DXR), the second enzyme of this metabolic route. D10 binds in vitro to recombinant DXR from P. falciparum and Escherichia coli, showing at 10 µM a ca. 50% inhibition of the bacterial enzyme. In silico docking analysis indicates that D10 associates with DXR in solvent-exposed regions outside the active center pocket. According to fluorescence confocal microscopy data, this aptamer specifically targets in P. falciparum in vitro cultures the apicoplast organelle where the MEP pathway is localized and is, therefore, a highly specific marker of red blood cells parasitized by Plasmodium vs. naïve erythrocytes. D10 is also selective for the detection of MEP+ bacteria (e.g., E. coli and Pseudomonas aeruginosa) vs. those lacking DXR (e.g., Enterococcus faecalis). Based on these results, we discuss the potential of DNA aptamers in the development of ligands that can outcompete the performance of the well-established antibody technology for future therapeutic and diagnostic approaches. MDPI 2022-11-19 /pmc/articles/PMC9695703/ /pubmed/36432706 http://dx.doi.org/10.3390/pharmaceutics14112515 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Roca, Carlota
Avalos-Padilla, Yunuen
Prieto-Simón, Beatriz
Iglesias, Valentín
Ramírez, Miriam
Imperial, Santiago
Fernàndez-Busquets, Xavier
Selection of an Aptamer against the Enzyme 1-deoxy-D-xylulose-5-phosphate Reductoisomerase from Plasmodium falciparum
title Selection of an Aptamer against the Enzyme 1-deoxy-D-xylulose-5-phosphate Reductoisomerase from Plasmodium falciparum
title_full Selection of an Aptamer against the Enzyme 1-deoxy-D-xylulose-5-phosphate Reductoisomerase from Plasmodium falciparum
title_fullStr Selection of an Aptamer against the Enzyme 1-deoxy-D-xylulose-5-phosphate Reductoisomerase from Plasmodium falciparum
title_full_unstemmed Selection of an Aptamer against the Enzyme 1-deoxy-D-xylulose-5-phosphate Reductoisomerase from Plasmodium falciparum
title_short Selection of an Aptamer against the Enzyme 1-deoxy-D-xylulose-5-phosphate Reductoisomerase from Plasmodium falciparum
title_sort selection of an aptamer against the enzyme 1-deoxy-d-xylulose-5-phosphate reductoisomerase from plasmodium falciparum
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9695703/
https://www.ncbi.nlm.nih.gov/pubmed/36432706
http://dx.doi.org/10.3390/pharmaceutics14112515
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