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Targeting the fungal plasma membrane proton pump
B C Monk1, A B Mason, T B Kardos
1Department of Oral Biology and Oral Pathology, University of Otago, Dunedin, New Zealand.
Acta Biochimica Polonica
|January 1, 1995
Summary
Developing new antifungal agents requires targeting the fungal proton pump, an essential enzyme. Inhibiting this plasma membrane ATPase, particularly with chimeric constructs, proves fungicidal and offers a novel therapeutic strategy against fungal infections.
Area of Science:
- Biochemistry
- Molecular Biology
- Antifungal Drug Discovery
Background:
- The plasma membrane proton pumping ATPase is a crucial enzyme in fungi like Saccharomyces cerevisiae.
- This enzyme belongs to the P-type ATPase family, which includes therapeutically targeted enzymes in humans.
- Existing drugs targeting similar ATPases highlight their potential as antifungal targets.
Purpose of the Study:
- To explore the fungal proton pump's membrane sector and ectodomain as a novel antifungal target.
- To investigate the role of specific transmembrane loops in fungal ATPase activity and regulation.
- To develop a basis for screening inhibitors against opportunistic fungal pathogens.
Main Methods:
- Constructing and expressing chimeric ATPases in S. cerevisiae using components from Candida albicans.
- Utilizing acid-activated omeprazole to assess the fungicidal effects of ATPase inhibition.
- Generating mutant panels with altered ATPase regulation and cell surface cysteine residues.
Main Results:
- Extensive inhibition of the S. cerevisiae ATPase was demonstrated to be fungicidal.
- Transmembrane loops 1+2 and 3+4 from Candida albicans function as a distinct domain within the chimeric ATPase.
- A model of the ATPase membrane sector and ectodomain was developed, integrating experimental data.
Conclusions:
- The fungal proton pump's membrane sector and ectodomain represent a promising target for broad-spectrum antifungal agents.
- Chimeric ATPases incorporating fungal pathogen sequences provide a valuable platform for drug discovery and development.
- Targeting this essential enzyme offers a new mechanistic class for combating fungal infections.