Gil-Navarro I, Gil ML, Cervera AM, Casanova M, Martinez JP, Gozalbo D; American Society for Microbiology. General Meeting.
Abstr Gen Meet Am Soc Microbiol. 1997 May 4-8; 97: 268 (abstract no. F51).
School of Pharmacy, University of Valencia, Spain.
A deltagt11 cDNA library from Candida albicans ATCC 26555 was screened using pooled sera from two patients with systemic candidiasis and five neutropenic patients with a high level of IgM antibodies anti-C. albicans. Seven clones were isolated from 60,000 recombinant phages. The most reactive one contained a 0.9 kb cDNA encoding a polypeptide immunoreactive only with sera from patients with systemic candidiasis. The whole gene was isolated using this cDNA as a probe. The nucleotide sequence of the coding region showed homology (76-78%) to the Saccharomyces cerevisiae TDH1 to TDH3 genes coding for glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and their amino acid sequences showed 76% identity; thus, this gene has been denominated CaTDH1. A rabbit polyclonal antiserum against the purified cytosolic C. albicans GAPDH (PAb anti-CA-GAPDH) was used to identify the GAPDH in the beta-mercaptoethanol extracts containing cell wall moieties. Indirect immunofluorescence and semiquantitative flow cytometry analysis demonstrated the presence of GAPDH at the C. albicans cell surface, particularly on blasto-conidial cells. In addition, a dose dependent GAPDH enzymatic activity was detected in intact blastoconidia and germ tube cells, which was reduced by pretreatment of the cells by trypsin, formaldehyde and PAb anti-CA-GAPDH. These observations indicate that an immunogenic, enzymatically active cell wall-associated form of GAPDH is found at the cell surface of C. albicans.
Publication Types:
Keywords:
- Amino Acid Sequence
- Animals
- Antigens, Fungal
- Antigens, Surface
- Base Sequence
- Candida albicans
- Candidiasis
- Cell Wall
- DNA, Complementary
- Fluorescent Antibody Technique, Indirect
- Glyceraldehyde-3-Phosphate Dehydrogenases
- Humans
- Phosphoglycerate Kinase
- Rabbits
- Saccharomyces cerevisiae
- Saccharomyces cerevisiae Proteins
- enzymology
- genetics
- immunology
Other ID:
UI: 102235353
From Meeting Abstracts