Identification of Host Receptors for Fungi Using Whole Cell Affinity Purification

Methods Mol Biol. 2021:2260:27-36. doi: 10.1007/978-1-0716-1182-1_3.

Abstract

Receptors on endothelial and epithelial cells often recognize molecules that are expressed by fungi, and only a limited number of these receptors have been identified to date. Here, we describe a method for identifying novel host cell receptors for fungi that uses intact organisms to precipitate biotin-labelled host cell membrane proteins, which are then detected by immunoblotting with an anti-biotin antibody. Presented here is the method to use for identification of membrane proteins that bind to C. albicans.

Keywords: Endothelial cells; Epithelial cells; Fungi; Immunoblotting; Invasion; Receptor.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Biotinylation
  • Blotting, Western*
  • Candida albicans / metabolism*
  • Cells, Cultured
  • Centrifugation
  • Endothelial Cells / metabolism*
  • Endothelial Cells / microbiology
  • Epithelial Cells / metabolism*
  • Epithelial Cells / microbiology
  • Fungal Proteins / metabolism*
  • Host-Pathogen Interactions
  • Humans
  • Protein Binding
  • Receptors, Cell Surface / isolation & purification*

Substances

  • Fungal Proteins
  • Receptors, Cell Surface