Towards the Toxoplasma gondii proteome: position of 13 parasite excretory antigens on a standardized map of two-dimensionally separated tachyzoite proteins.
High resolution two-dimensional separation of Toxoplasma gondii tachyzoite lysate revealed up to 224 distinct protein spots in Coomassie-stained gel. Computional matching of 14 digitized gels yielded a standard two-dimensional proteome map. The excretory T. gondii dense granule proteins GRA1-GRA8, S16/acid phosphatase, nucleoside triphosphate hydrolase, and H4 were identified by Western blotting of both total gel and isolated protein spots. In addition, two excretory antigens defined by parasite-specific monoclonal T cells, p36 and p40, were mapped by a novel T-cell blotting technique based on electroeluting single protein spots and testing the eluates for antigenic activity against the T-cell clones. In summary, these results represent a first step in Toxoplasma proteome analysis.
Dlugonska
H
H
Institute of Microbiology and Immunology, University of Lodz, Poland.
Dytnerska
K
K
Reichmann
G
G
Stachelhaus
S
S
Fischer
H G
HG
D000818:Animals; D000953:Antigens, Protozoan / Q000032:analysis* / Q000276:immunology; D015153:Blotting, Western; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology; D015180:Electrophoresis, Gel, Two-Dimensional; D008213:Lymphocyte Activation; D010449:Peptide Mapping / Q000379:methods; D020543:Proteome; D015800:Protozoan Proteins / Q000032:analysis; D014122:Toxoplasma / Q000737:chemistry* / Q000276:immunology
D000953:Antigens, Protozoan; D020543:Proteome; D015800:Protozoan Proteins; C408624:excretory-secretory antigen, Toxoplasma