Skip to main content
PMC home page
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1988 Nov;26(11):2250–2256. doi: 10.1128/jcm.26.11.2250-2256.1988

Characterization of a soluble protein of Coccidiodes immitis with activity as an immunodiffusion-complement fixation antigen.

B L Zimmer 1, D Pappagianis 1
PMCID: PMC266870  PMID: 3148631

Abstract

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.

Full text

PDF
2253

Images in this article

2252Image
on p.2252
2252Image
on p.2252
2252Image
on p.2252
2253Image
on p.2253
2253Image
on p.2253
2254Image
on p.2254

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Anderson K. L., Wheat R. W., Conant N. F. Fractionation and composition studies of skin test-active components of sensitins from Coccidioides immitis. Appl Microbiol. 1971 Sep;22(3):294–299. doi: 10.1128/am.22.3.294-299.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. BERMAN R. J., FRIEDMAN L., ROESSLER W. G., SMITH C. E. The virulence and infectivity of twenty-seven strains of Coccidioides immitis. Am J Hyg. 1956 Sep;64(2):198–210. doi: 10.1093/oxfordjournals.aje.a119834. [DOI] [PubMed] [Google Scholar]
  3. CANFIELD R. E., LIU A. K. THE DISULFIDE BONDS OF EGG WHITE LYSOZYME (MURAMIDASE). J Biol Chem. 1965 May;240:1997–2002. [PubMed] [Google Scholar]
  4. Calhoun D. L., Osir E. O., Dugger K. O., Galgiani J. N., Law J. H. Humoral antibody responses to specific antigens of Coccidioides immitis. J Infect Dis. 1986 Aug;154(2):265–272. doi: 10.1093/infdis/154.2.265. [DOI] [PubMed] [Google Scholar]
  5. Cox R. A., Britt L. A. Antigenic identity of biologically active antigens in coccidioidin and spherulin. Infect Immun. 1987 Nov;55(11):2590–2596. doi: 10.1128/iai.55.11.2590-2596.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cox R. A., Britt L. A., Michael R. A. Isolation of Coccidioides immitis F antigen by immunoaffinity chromatography with monospecific antiserum. Infect Immun. 1987 Jan;55(1):227–232. doi: 10.1128/iai.55.1.227-232.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Huppert M., Spratt N. S., Vukovich K. R., Sun S. H., Rice E. H. Antigenic analysis of coccidioidin and spherulin determined by two-dimensional immunoelectrophoresis. Infect Immun. 1978 May;20(2):541–551. doi: 10.1128/iai.20.2.541-551.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Ibrahim A. B., Pappagianis D. Experimental induction of anergy to coccidioidin by antigens of Coccidioides immitis. Infect Immun. 1973 May;7(5):786–794. doi: 10.1128/iai.7.5.786-794.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. LEVINE H. B., COBB J. M., SMITH C. E. Immunity to coccidioi-domycosis induced in mice by purified spherule, arthrospore, and mycelial vaccines. Trans N Y Acad Sci. 1960 Apr;22:436–449. doi: 10.1111/j.2164-0947.1960.tb00711.x. [DOI] [PubMed] [Google Scholar]
  10. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  11. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  12. Levine H. B., Pappagianis D., Cobb J. M. Development of vaccines for coccidioidomycosis. Mycopathol Mycol Appl. 1970;41(1):177–185. doi: 10.1007/BF02051493. [DOI] [PubMed] [Google Scholar]
  13. Merril C. R., Goldman D., Sedman S. A., Ebert M. H. Ultrasensitive stain for proteins in polyacrylamide gels shows regional variation in cerebrospinal fluid proteins. Science. 1981 Mar 27;211(4489):1437–1438. doi: 10.1126/science.6162199. [DOI] [PubMed] [Google Scholar]
  14. Naot Y., Guptill D. R., Mullenax J., Remington J. S. Characterization of Toxoplasma gondii antigens that react with human immunoglobulin M and immunoglobulin G antibodies. Infect Immun. 1983 Jul;41(1):331–338. doi: 10.1128/iai.41.1.331-338.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Osawa T., Tsuji T. Fractionation and structural assessment of oligosaccharides and glycopeptides by use of immobilized lectins. Annu Rev Biochem. 1987;56:21–42. doi: 10.1146/annurev.bi.56.070187.000321. [DOI] [PubMed] [Google Scholar]
  16. PAPPAGIANIS D., LINDSEY N. J., SMITH C. E., SAITO M. T. ANTIBODIES IN HUMAN COCCIDIOIDOMYCOSIS: IMMUNOELECTROPHORETIC PROPERTIES. Proc Soc Exp Biol Med. 1965 Jan;118:118–122. doi: 10.3181/00379727-118-29773. [DOI] [PubMed] [Google Scholar]
  17. PAPPAGIANIS D., PUTMAN E. W., KOBAYASHI G. S. Polysaccharide of Coccidioides immitis. J Bacteriol. 1961 Nov;82:714–723. doi: 10.1128/jb.82.5.714-723.1961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Resnick S., Pappagianis D., McKerrow J. H. Proteinase production by the parasitic cycle of the pathogenic fungus Coccidioides immitis. Infect Immun. 1987 Nov;55(11):2807–2815. doi: 10.1128/iai.55.11.2807-2815.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. SMITH C. E., SAITO M. T., SIMONS S. A. Pattern of 39,500 serologic tests in coccidioidomycosis. J Am Med Assoc. 1956 Feb 18;160(7):546–552. doi: 10.1001/jama.1956.02960420026008. [DOI] [PubMed] [Google Scholar]
  20. Sawaki Y., Huppert M., Bailey J. W., Yagi Y. Patterns of human antibody reactions in coccidioidomycosis. J Bacteriol. 1966 Jan;91(1):422–427. doi: 10.1128/jb.91.1.422-427.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Scheer E., Terai T., Kulkarni S., Conant N. F., Wheat R. W., Lowe E. P. Unusual reducing sugar from Coccidioides immitis. J Bacteriol. 1970 Aug;103(2):525–526. doi: 10.1128/jb.103.2.525-526.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Yuan L., Cole G. T. Isolation and characterization of an extracellular proteinase of Coccidioides immitis. Infect Immun. 1987 Sep;55(9):1970–1978. doi: 10.1128/iai.55.9.1970-1978.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Zimmer B. L., Pappagianis D. Comparison of immunoblot analyses of spherule-endospore-phase extracellular protein and mycelial-phase antigen of Coccidioides immitis. Infect Immun. 1986 Jul;53(1):64–70. doi: 10.1128/iai.53.1.64-70.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Clinical Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES