Complications in cell-surface labelling by biotinylation of Candida albicans due to avidin conjugate binding to cell-wall proteins.

Published

Journal Article

Initial contact between the opportunistic fungal pathogen Candida albicans and host tissue occurs at the cell surface. Biotin derivatives have been used to label the cell-surface proteins of yeasts, with labelled proteins subsequently detected by avidin-reporter conjugates. Previous work has indicated that avidin can bind to C. albicans proteins in the absence of biotin, suggesting a possible host-cell-recognition mechanism by fungal cell-surface proteins. To investigate this mechanism, Western blots of proteins extracted from biotinylated and mock-treated cells were probed with avidin or modified-avidin reagents. Each avidin reagent bound to cell-wall proteins extracted from non-biotinylated cells. Binding did not appear to be due to the lectin-like activity of the cell-wall proteins of C. albicans or to the presence of biotin in the sample itself. Binding was inhibited by added biotin, by the chaotrope KSCN and by NaCl in a concentration-dependent manner, although inhibition varied among the avidin conjugates tested. Thus, the non-specific binding of avidin to the cell-wall proteins of C. albicans appears to involve hydrophobic and electrostatic interactions, depending on the particular avidin species. These observations demonstrate potential pitfalls in the use of avidin-biotin complexes to identify cell-surface molecules and could provide insights into protein-protein interactions at the C. albicans cell wall.

Full Text

Duke Authors

Cited Authors

  • Masuoka, J; Guthrie, LN; Hazen, KC

Published Date

  • April 2002

Published In

Volume / Issue

  • 148 / Pt 4

Start / End Page

  • 1073 - 1079

PubMed ID

  • 11932452

Pubmed Central ID

  • 11932452

International Standard Serial Number (ISSN)

  • 1350-0872

Digital Object Identifier (DOI)

  • 10.1099/00221287-148-4-1073

Language

  • eng

Conference Location

  • England