A luminescence assay for natural product inhibitors of the Mycobacterium tuberculosis proteasome
| Autor: | John R. Porter, Amber Gunderwala |
|---|---|
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Fluorescence assay Plant Science Biochemistry Analytical Chemistry law.invention Mycobacterium tuberculosis 03 medical and health sciences chemistry.chemical_compound law Drug Discovery Chemiluminescence Natural product biology Assay General Medicine biology.organism_classification Molecular biology 030104 developmental biology Proteasome activity Complementary and alternative medicine chemistry Proteasome Molecular Medicine Luminescence Food Science |
| Zdroj: | Phytochemical Analysis. 27:126-132 |
| ISSN: | 0958-0344 |
| DOI: | 10.1002/pca.2607 |
| Popis: | Introduction Mycobacterium tuberculosis (Mtb) causes a large global burden of disease, with a high mortality rate in healthy and immuno-compromised patients. A number of molecular targets have been identified for treatment of this disease, including the Mtb proteasome. The Mtb proteasome enhances Mtb survival during nitrosative and oxidative stress in the latent, non-replicative phase. Therefore, Mtb proteasome inhibition could help to combat Mtb infections that do not respond to current therapies. Objective To develop and validate a novel biochemical assay to assess Mtb proteasome activity in the presence of organic and aqueous plant test extracts. Method Fluorescence (photoluminescence) and luminescence (chemiluminescence) assays were investigated as potential methods to determine the robustness and repeatability for use in screening natural product extracts for Mtb proteasome inhibitors. Results The fluorescence assay, used widely for Mtb proteasome activity assays, was subject to interference due to the natural fluorescence of compounds in many of the extracts; there is little interference with the luminescence approach. As proof of principle, we used the luminescence assay to screen a small set of plant test extracts. Conclusions Luminescence is the more suitable assay for assay of plant natural product extracts. The sensitivities of the luminescence and fluorescence assays are comparable. A Z′-factor of 0.58 for the luminescence assay makes it suitable for medium-to-high throughput screening efforts. Copyright © 2016 John Wiley & Sons, Ltd. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Plný text