Time-Resolved Fluorescence Assay for Measuring Oxygen Consumption Rates in Staphylococcus Aureus

Overview

Journal Methods Mol Biol

Specialty Molecular Biology

Date 2021 Jul 15

PMID 34264464

Authors

Logan L Bulock

Jennifer L Endres

Marat R Sadykov

Affiliations

Soon will be listed here.

Abstract

Oxygen consumption is a fundamental characteristic of staphylococcal physiology reflecting the energy and metabolic state of the bacterial cell. During aerobic growth, oxygen consumption rates (OCR) depend on nutrient availability and vary at different growth stages. The measurement of oxygen consumption rates provides a versatile tool to characterize the impact of various mutations, environmental cues, and antibiotics on bacterial growth and fitness. In this chapter, we describe a MitoXpress Xtra-based oxygen consumption assay for fast and reliable determination of respiration rates in Staphylococcus aureus. This highly reproducible and simple method requires a minimal set of reagents and allows rapid screening of multiple samples through real-time determination of the OCR with an oxygen-sensing probe and fluorescence plate reader.

References

Balemans W, Vranckx L, Lounis N, Pop O, Guillemont J, Vergauwen K . Novel antibiotics targeting respiratory ATP synthesis in Gram-positive pathogenic bacteria. Antimicrob Agents Chemother. 2012; 56(8):4131-9. PMC: 3421580. DOI: 10.1128/AAC.00273-12. View

Dibrov P, Dibrov E, Maddaford T, Kenneth M, Nelson J, Resch C . Development of a novel rationally designed antibiotic to inhibit a nontraditional bacterial target. Can J Physiol Pharmacol. 2017; 95(5):595-603. DOI: 10.1139/cjpp-2016-0505. View

Halsey C, Lei S, Wax J, Lehman M, Nuxoll A, Steinke L . Amino Acid Catabolism in and the Function of Carbon Catabolite Repression. mBio. 2017; 8(1). PMC: 5312079. DOI: 10.1128/mBio.01434-16. View

Sadykov M, Ahn J, Widhelm T, Eckrich V, Endres J, Driks A . Poly(3-hydroxybutyrate) fuels the tricarboxylic acid cycle and de novo lipid biosynthesis during Bacillus anthracis sporulation. Mol Microbiol. 2017; 104(5):793-803. DOI: 10.1111/mmi.13665. View

Jasionek G, Ogurtsov V, Papkovsky D . Rapid detection and respirometric profiling of aerobic bacteria on panels of selective media. J Appl Microbiol. 2012; 114(2):423-32. DOI: 10.1111/jam.12049. View