Transport across Membranes: Techniques for Measuring Efflux in Fungal Cells

Methods Mol Biol. 2023:2658:201-213. doi: 10.1007/978-1-0716-3155-3_14.

Abstract

One of the most prevalent mechanisms of antifungal drug resistance is export of the molecule from the fungal cells through the action of putative efflux pumps or transporters. Drug efflux is a particularly common mechanism of resistance to azole antifungals, one of the most widely used classes of antifungal drugs. Here, we provide detailed protocols for two assays of small-molecule efflux activity: rhodamine 6G efflux and alanine-naphthylamide accumulation. Protocols applicable to both yeast and filamentous fungi are provided.

Keywords: ATP-binding cassette transporters; Ala-Nap; Efflux; Major facilitator transporters; Membrane transporters; Rhodamine 6G.

MeSH terms

  • ATP-Binding Cassette Transporters* / metabolism
  • Antifungal Agents / pharmacology
  • Biological Transport
  • Candida albicans / metabolism
  • Drug Resistance, Fungal
  • Fungal Proteins* / metabolism
  • Membrane Transport Proteins / metabolism
  • Saccharomyces cerevisiae / metabolism

Substances

  • ATP-Binding Cassette Transporters
  • Fungal Proteins
  • Membrane Transport Proteins
  • Antifungal Agents