Characterization of a soluble protein of Coccidiodes immitis with activity as an immunodiffusion-complement fixation antigen.
AUTOR(ES)
Zimmer, B L
RESUMO
A 48-kilodalton (kDa) electrophoretically distinct antigen from Coccidioides immitis mycelial- and spherule-endospore-phase filtrates was previously associated by immunoblotting with the immunodiffusion band that corresponds to complement-fixing activity (ID-CF). To characterize this antigen and its precursor, both mycelial- and spherule-endospore-phase filtrates were fractionated by size exclusion chromatography, lectin affinity chromatography, and nondenaturing electrophoresis. By size exclusion chromatography, most of the protein and carbohydrate of the crude filtrates eluted in a peak of average molecular size less than 30 kDa, although other components were detected. ID-CF activity was associated with the component at a relative mobility of 110 kDa. Fractions containing the ID band that corresponded to tube precipitin activity occurred from 200 to 40 kDa. The appearance of the 48-kDa band in denaturing sodium dodecyl sulfate-polyacrylamide gel electrophoresis (PAGE) specifically coincided with the fractions containing ID-CF activity. Nondenaturing PAGE of filtrates showed silver-stainable and immunoblot-reactive bands in the region of 110 kDa. Prior treatment with pronase destroyed this electrophoretically separable antigen, whereas periodate had no effect. Trypsin did not affect the 110-kDa band in unheated or unreduced antigen. Mycelial filtrates were chromatographed on lentil lectin or concanavalin A-Sepharose 4B to deplete them of glucose- or mannose-containing carbohydrate. The effluent fraction contained ID-CF activity and, upon denaturing electrophoresis, the 48-kDa antigen. The 110-kDa protein represents the ID-CF antigen which is heat labile and denatured to a 48-kDa band by sodium dodecyl sulfate-PAGE.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=266870Documentos Relacionados
- The coccidioidal complement fixation and immunodiffusion-complement fixation antigen is a chitinase.
- Molecular cloning and characterization of the Coccidioides immitis complement fixation/chitinase antigen.
- Amino-terminal sequence analysis of the Coccidioides immitis chitinase/immunodiffusion-complement fixation protein.
- Partial characterization of the principal soluble antigens associated with the coronavirus of transmissible gastroenteritis by complement fixation and immunodiffusion.
- Production and characterization of a monoclonal antibody to the complement fixation antigen of Coccidioides immitis.