A catalytic bioscavenger with improved stability and reduced susceptibility to oxidation for treatment of acute poisoning with neurotoxic organophosphorus compounds
| Autor: | Laura Job, Anja Köhler, Franz Worek, Benjamin Escher, Arne Skerra |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Protein Denaturation Hot Temperature Antidotes Soman Toxicology medicine.disease_cause 03 medical and health sciences Organophosphate Poisoning Organophosphorus Compounds 0302 clinical medicine Bacterial Proteins Drug Stability Enzyme Stability medicine Brevundimonas diminuta Escherichia coli Nerve agent chemistry.chemical_classification Organothiophosphorus Compounds Caulobacteraceae General Medicine Sarin Recombinant Proteins Acute toxicity Amino acid Phosphoric Triester Hydrolases 030104 developmental biology Enzyme Biochemistry chemistry Mutation Nerve Agents Oxidation-Reduction 030217 neurology & neurosurgery Oxidative stress Cysteine medicine.drug |
| Zdroj: | Toxicology Letters. 321:138-145 |
| ISSN: | 0378-4274 |
| DOI: | 10.1016/j.toxlet.2019.12.030 |
| Popis: | Organophosphorus (OP) 1 nerve agents pose a severe toxicological threat, both after dissemination in military conflicts and by terrorists. Hydrolytic enzymes, which may be administered into the blood stream of victims by injection and can decompose the circulating nerve agent into non-toxic metabolites in vivo, could offer a treatment. Indeed, for the phosphotriesterase found in the bacterium Brevundimonas diminuta (BdPTE), 2 engineered versions with improved catalytic efficiencies have been described; yet, their biochemical stabilities are insufficient for therapeutic use. Here, we describe the application of rational protein design to develop novel mutants of BdPTE that are less susceptible to oxidative damage. In particular, the replacement of two unpaired cysteine residues by more inert amino acids led to higher stability while maintaining high catalytic activity towards a broad spectrum of substrates, including OP pesticides and V-type nerve agents. The mutant BdPTE enzymes were produced in Escherichia coli, purified to homogeneity, and their biochemical and enzymological properties were assessed. Several candidates both revealed enhanced thermal stability and were less susceptible to oxidative stress, as demonstrated by mass spectrometry. These mutants of BdPTE may show promise for the treatment of acute intoxications by nerve agents as well as OP pesticides. |
| Databáze: | OpenAIRE |
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