Kinetic characterisation and inhibitor sensitivity of Candida albicans and Candida auris recombinant AOX expressed in a self-assembled proteoliposome system

Copsey, Alice C, Barsottini, Mario R O, May, Benjamin, Xu, Fei, Albury, Mary S, Young, Luke and Moore, Anthony L (2021) Kinetic characterisation and inhibitor sensitivity of Candida albicans and Candida auris recombinant AOX expressed in a self-assembled proteoliposome system. Scientific Reports, 11. a14748 1-10. ISSN 2045-2322

[img] PDF - Published Version
Available under License Creative Commons Attribution.

Download (2MB)

Abstract

Candidemia caused by Candida spp. is a serious threat in hospital settings being a major cause of acquired infection and death and a possible contributor to Covid-19 mortality. Candidemia incidence has been rising worldwide following increases in fungicide-resistant pathogens highlighting the need for more effective antifungal agents with novel modes of action. The membrane-bound enzyme alternative oxidase (AOX) promotes fungicide resistance and is absent in humans making it a desirable therapeutic target. However, the lipophilic nature of the AOX substrate (ubiquinol-10) has hindered its kinetic characterisation in physiologically-relevant conditions. Here, we present the purification and expression of recombinant AOXs from C. albicans and C. auris in a self-assembled proteoliposome (PL) system. Kinetic parameters (Km and Vmax) with respect to ubiquinol-10 have been determined. The PL system has also been employed in dose–response assays with novel AOX inhibitors. Such information is critical for the future development of novel treatments for Candidemia.

Item Type: Article
Keywords: Antifungal Agents, Candida albicans, Drug Resistance, Fungal, Enzyme Inhibitors, Fungal Proteins, Kinetics, Liposomes, Mitochondrial Proteins, Oxidoreductases, Plant Proteins, Recombinant Proteins
Schools and Departments: School of Life Sciences > Biochemistry
SWORD Depositor: Mx Elements Account
Depositing User: Mx Elements Account
Date Deposited: 09 Sep 2021 11:13
Last Modified: 09 Sep 2021 11:15
URI: http://sro.sussex.ac.uk/id/eprint/101598

View download statistics for this item

📧 Request an update