Analysis of cytoplasmic antigens of the yeast and mycelial phases of Candida albicans by two-dimensional electrophoresis.

Published

Journal Article

The extent of the macromolecular change accompanying yeast to mycelium morphogenesis of Candida albicans was analyzed by two-dimensional gel electrophoresis of the cytoplasmic proteins of the two growth forms after antibody cross-absorption experiments. Pure cultures of yeasts and true hyphae (i.e., without concomitant production of pseudohyphae) were grown in a synthetic low-sulfate medium (LSM). The two strains selected for this study were strain 4918, which produces pure mycelial (M) cultures in LSM at 37 degrees C (designated 4918-37M) and yeasts (Y) at 24 degrees C (4918-24Y), and strain 2252, which produces yeasts exclusively at both 24 and 37 degrees C in LSM (2252-24Y and 2252-37Y). The proteins of both strains were labeled at both temperatures with [35S]sulfate, and cytoplasmic fractions were prepared by mechanical disruption and ultracentrifugation. Rabbits were immunized with the 4918-24Y and 4918-37M cytoplasmic fractions to produce anti-yeast-phase and anti-mycelial-phase hyperimmune sera. Each radiolabeled cytoplasmic fraction was absorbed with anti-mycelial-phase immunoglobulin, anti-yeast immunoglobulin, and immunoglobulin from normal rabbit serum. Staphyloccal protein A was used to remove immune complexes. The labeled, nonabsorbed proteins were also analyzed by two-dimensional electrophoresis. Highly reproducible protein spot patterns were obtained which defined hundreds of proteins in each extract. The specificity of the immunoglobulin hundreds of proteins in each extract. The specificity of the immunoglobulin preparations was extremely broad, and as many as 168 cytoplasmic antigens were detected. Eighty-three antigens were recognized in the mycelial-phase extract only by the anti-mycelial-phase immunoglobulin. However, comparative analysis revealed that all of these proteins were present in at least one other extract. Therefore, none of them was unique to the mycelial morphology. Eleven antigens were detected in the 2252-37Y extract that were not present in the extracts from strain 4918, which indicates that proteins obtained from different strains may express similar antigenic determinants, but differ in their physiochemical properties.

Full Text

Duke Authors

Cited Authors

  • Manning, M; Mitchell, TG

Published Date

  • November 1, 1980

Published In

Volume / Issue

  • 30 / 2

Start / End Page

  • 484 - 495

PubMed ID

  • 7002799

Pubmed Central ID

  • 7002799

International Standard Serial Number (ISSN)

  • 0019-9567

Language

  • eng

Conference Location

  • United States