Synthesis and interaction of terminal unsaturated chemical probes with Mycobacterium tuberculosis CYP124A1
| Autor: | Luz Díaz-Storani, Jonathan B. Johnston, Andrea B. J. Bracca, Diego M. Moreno, Exequiel O. J. Porta, Anaelle A. Clary, María Sol Ballari, Guillermo R. Labadie |
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| Rok vydání: | 2021 |
| Předmět: |
Stereochemistry
Clinical Biochemistry Pharmaceutical Science Biochemistry Mycobacterium tuberculosis chemistry.chemical_compound Drug Discovery medicine Bound water Molecular Biology Heme chemistry.chemical_classification Molecular Structure biology Terpenes Chemistry Organic Chemistry Active site Farnesol biology.organism_classification Enzyme Docking (molecular) Molecular Probes biology.protein Molecular Medicine Ferric lipids (amino acids peptides and proteins) Cytochrome P-450 CYP4A medicine.drug |
| Zdroj: | Bioorganic & Medicinal Chemistry. 44:116304 |
| ISSN: | 0968-0896 |
| DOI: | 10.1016/j.bmc.2021.116304 |
| Popis: | A series of C15-C20 isoprenyl derivatives bearing terminal alkenyl and alkynyl groups were synthesized as possible substrates of the methyl-branched lipid ω-hydroxylase CYP124A1 from Mycobacterium tuberculosis. The interactions of each compound with the enzyme active site were characterized using UV-vis spectroscopy. We found that C10 and C15 analogs bind with similar affinity to the corresponding parent C10 and C15 substrates geraniol and farnesol, respectively. Three analogs (C10-ω-ene, C10-ω-yne, C15-ω-yne) interact with the proximal side of the heme iron by coordinating to the oxygen atom of the ferric heme, as judged by the appearance of typical Type-IA binding spectra. On the other hand, the C15-ω-ene analog interacts with the ferric heme by displacing the bound water that generates a typical Type I binding spectrum. We were unable to detect P450-mediated oxidation of these probes following extended incubations with CYP124A1 in our reconstituted assay system, whereas a control reaction containing farnesol was converted to ω-hydroxy farnesol under the same conditions. To understand the lack of detectable oxidation, we explored the possibility that the analogs were acting as mechanism-based inhibitors, but we were unable to detect time-dependent loss of enzymatic activity. In order to gain insight into the lack of detectable turnover or time-dependent inhibition, we examined the interaction of each compound with the CYP124A1 active site using molecular docking simulations. The docking studies revealed a binding mode where the terminal unsaturated functional groups were sequestered within the methyl-binding pocket, rather than positioned close to the heme iron for oxidation. These results aid in the design of specific inhibitors of Mtb-CYP124A1, an interesting enzyme that is implicated in the oxidation of methyl-branched lipids, including cholesterol, within a deadly human pathogen. |
| Databáze: | OpenAIRE |
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