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A Simple Assay for Inhibitors of Mycobacterial Oxidative Phosphorylation by Serena A. Harden; Gautier M. Courbon; Yingke Liang; Angelina S. Kim; John L. Rubinstein is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.

What is A Simple Assay for Inhibitors of Mycobacterial Oxidative Phosphorylation about?

Oxidative phosphorylation, the combined activities of the electron transport chain (ETC) and ATP synthase, has emerged as a valuable target for antibiotics to treat infection with __Mycobacterium tuberculosis__ and related pathogens. In oxidative phosphorylation, the ETC establishes a transmembrane electrochemical proton gradient that powers ATP synthesis. Monitoring oxidative phosphorylation with luciferase-based detection of ATP synthesis or measurement of oxygen consumption can be technically challenging and expensive. These limitations reduce the utility of these methods for characterization of mycobacterial oxidative phosphorylation inhibitors. Here, we show that fluorescence-based measurement of acidification of inverted membrane vesicles (IMVs) can detect and distinguish between inhibition of the ETC, inhibition of ATP synthase, and nonspecific membrane uncoupling. In this assay, IMVs from __Mycobacterium smegmatis__ are acidified either through the activity of the ETC or ATP synthase, the latter modified genetically to allow it to serve as an ATP-driven proton pump. Acidification is monitored by fluorescence from 9-amino-6-chloro-2-methoxyacridine, which accumulates and que

Who reads A Simple Assay for Inhibitors of Mycobacterial Oxidative Phosphorylation?

Author

Serena A. Harden; Gautier M. Courbon; Yingke Liang; Angelina S. Kim; John L. Rubinstein

Publisher

American Society for Biochemistry and Molecular Biology

Published

2023

Language

EN

Field

Biochemistry, Genetics and Molecular Biology (Life Sciences)