DNA aptamers developed against a soman derivative cross-react with the methylphosphonic acid core but not with flanking hydrophobic groups
| Autor: | Douglas M. Cerasoli, David E. Lenz, C. Linn Cadieux, Taylor Phillips, John G. Bruno, Maria P. Carrillo |
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| Rok vydání: | 2009 |
| Předmět: |
Models
Molecular Molecular model Stereochemistry Aptamer Molecular Sequence Data Soman Cross Reactions chemistry.chemical_compound Organophosphorus Compounds Structural Biology Nucleotide Molecular Biology Methylphosphonic acid chemistry.chemical_classification Base Sequence SELEX Aptamer Technique Titrimetry Aptamers Nucleotide Phosphonate Spectrometry Fluorescence chemistry Nucleic Acid Conformation Sequence motif Hydrophobic and Hydrophilic Interactions Systematic evolution of ligands by exponential enrichment Conjugate |
| Zdroj: | Journal of Molecular Recognition. 22:197-204 |
| ISSN: | 1099-1352 0952-3499 |
| Popis: | Twelve rounds of systematic evolution of ligands by exponential enrichment (SELEX) were conducted against a magnetic bead conjugate of the para-aminophenylpinacolylmethylphosphonate (PAPMP) derivative of the organophosphorus (OP) nerve agent soman (GD). The goal was to develop DNA aptamers that could scavenge GD in vivo, thereby reducing or eliminating the toxic effects of this dangerous compound. Aptamers were sequenced and screened in peroxidase-based colorimetric plate assays after rounds 8 and 12 of SELEX. The aptamer candidate sequences exhibiting the highest affinity for the GD derivative from round 8 also reappeared in several clones from round 12. Each of the highest affinity PAPMP-binding aptamers also bound methylphosphonic acid (MPA). In addition, the aptamer with the highest overall affinity for PAPMP carried a sequence motif (TTTAGT) thought to bind MPA based on previously published data (J. Fluoresc 18: 867–876, 2008). This sequence motif was found in several other relatively high affinity PAPMP aptamer candidates as well. In studies with the nerve agent GD, pre-incubation of a large molar excess of aptamer candidates failed to protect human butyrylcholinesterase (BuChE) from inhibition. With the aid of three-dimensional molecular modeling of the GD derivative it appears that a hydrophilic cleft sandwiched between the pinacolyl group and the p-aminophenyl ring might channel nucleotide interactions to the phosphonate portion of the immobilized GD derivative. However, bona fide GD free in solution may be repulsed by the negative phosphate backbone of aptamers and rotate its phosphonate and fluorine moieties away from the aptamer to avoid being bound. Future attempts to develop aptamers to GD might benefit from immobilizing the pinacolyl group of bona fide GD to enhance exposure of the phosphonate and fluorine to the random DNA library. Copyright © 2008 John Wiley & Sons, Ltd. |
| Databáze: | OpenAIRE |
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