Skip to main content
PMC home page
Infection and Immunity logoLink to Infection and Immunity
. 1989 Jul;57(7):2149–2157. doi: 10.1128/iai.57.7.2149-2157.1989

Identification and isolation of a major cell surface glycoprotein of Pneumocystis carinii.

J A Radding 1, M Y Armstrong 1, E Ullu 1, F F Richards 1
PMCID: PMC313854  PMID: 2786505

Abstract

Radioiodination of rat-derived Pneumocystis carinii obtained from an in vitro culture demonstrated the presence of a major surface glycoprotein (gp120). The glycoprotein was of the high mannose type. It exhibited adherence properties similar to those observed in the intact organism. Under nonreducing conditions, it existed as an aggregate with a molecular weight in excess of 2 x 10(6). Surface aggregating behavior and adherent quality prevented isolation of the glycoprotein by conventional methods. The glycoprotein was purified by chromatography on hydroxyapatite in the presence of sodium dodecyl sulfate under reducing conditions.

Full text

PDF
2151

Images in this article

2152Image
on p.2152
2153Image
on p.2153
2153Image
on p.2153
2153Image
on p.2153
2154Image
on p.2154
2154Image
on p.2154
2155Image
on p.2155
2156Image
on p.2156

Selected References

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

  1. Armstrong M. Y., Richards F. F. Propagation and purification of rat Pneumocystis carinii in short-term cell culture. J Protozool. 1989 Jan-Feb;36(1):24S–27S. doi: 10.1111/j.1550-7408.1989.tb02677.x. [DOI] [PubMed] [Google Scholar]
  2. Bhavanandan V. P., Hegarty J. D. Identification of the mucin core protein by cell-free translation of messenger RNA from bovine submaxillary glands. J Biol Chem. 1987 Apr 25;262(12):5913–5917. [PubMed] [Google Scholar]
  3. Cushion M. T., Blase M. A., Walzer P. D. A method for isolation of RNA from Pneumocystis carinii. J Protozool. 1989 Jan-Feb;36(1):12S–14S. doi: 10.1111/j.1550-7408.1989.tb02668.x. [DOI] [PubMed] [Google Scholar]
  4. Cushion M. T., DeStefano J. A., Walzer P. D. Pneumocystis carinii: surface reactive carbohydrates detected by lectin probes. Exp Parasitol. 1988 Dec;67(2):137–147. doi: 10.1016/0014-4894(88)90061-6. [DOI] [PubMed] [Google Scholar]
  5. Cushion M. T., Walzer P. D. Growth and serial passage of Pneumocystis carinii in the A549 cell line. Infect Immun. 1984 May;44(2):245–251. doi: 10.1128/iai.44.2.245-251.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Edge A. S., Faltynek C. R., Hof L., Reichert L. E., Jr, Weber P. Deglycosylation of glycoproteins by trifluoromethanesulfonic acid. Anal Biochem. 1981 Nov 15;118(1):131–137. doi: 10.1016/0003-2697(81)90168-8. [DOI] [PubMed] [Google Scholar]
  7. Edman J. C., Kovacs J. A., Masur H., Santi D. V., Elwood H. J., Sogin M. L. Ribosomal RNA genes of Pneumocystis carinii. J Protozool. 1989 Jan-Feb;36(1):18S–20S. doi: 10.1111/j.1550-7408.1989.tb02672.x. [DOI] [PubMed] [Google Scholar]
  8. Elder J. H., Pickett R. A., 2nd, Hampton J., Lerner R. A. Radioiodination of proteins in single polyacrylamide gel slices. Tryptic peptide analysis of all the major members of complex multicomponent systems using microgram quantities of total protein. J Biol Chem. 1977 Sep 25;252(18):6510–6515. [PubMed] [Google Scholar]
  9. Fahim R. E., Specian R. D., Forstner G. G., Forstner J. F. Characterization and localization of the putative 'link' component in rat small-intestinal mucin. Biochem J. 1987 May 1;243(3):631–640. doi: 10.1042/bj2430631. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Fishman J. A., Ullu E., Armstrong M. Y., Richards F. F. Organization of DNA and RNA from rat Pneumocystis carinii. J Protozool. 1989 Jan-Feb;36(1):4S–5S. doi: 10.1111/j.1550-7408.1989.tb02663.x. [DOI] [PubMed] [Google Scholar]
  11. Gigliotti F., Stokes D. C., Cheatham A. B., Davis D. S., Hughes W. T. Development of murine monoclonal antibodies to Pneumocystis carinii. J Infect Dis. 1986 Aug;154(2):315–322. doi: 10.1093/infdis/154.2.315. [DOI] [PubMed] [Google Scholar]
  12. Graves D. C., McNabb S. J., Worley M. A., Downs T. D., Ivey M. H. Analyses of rat Pneumocystis carinii antigens recognized by human and rat antibodies by using western immunoblotting. Infect Immun. 1986 Oct;54(1):96–103. doi: 10.1128/iai.54.1.96-103.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hill H. D., Jr, Reynolds J. A., Hill R. L. Purification, composition, molecular weight, and subunit structure of ovine submaxillary mucin. J Biol Chem. 1977 Jun 10;252(11):3791–3798. [PubMed] [Google Scholar]
  14. Howard R. J., Kaushal D. C., Carter R. Radioiodination of parasite antigens with 1,3,4,6-tetrachloro-3 alpha, 6 alpha-diphenylglycoluril (IODOGEN): studies with zygotes of Plasmodium gallinaceum. J Protozool. 1982 Feb;29(1):114–117. doi: 10.1111/j.1550-7408.1982.tb02891.x. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. Moss B., Rosenblum E. N. Hydroxylapatite chromatography of protein-sodium dodecyl sulfate complexes. A new method for the separation of polypeptide subunits. J Biol Chem. 1972 Aug 25;247(16):5194–5198. [PubMed] [Google Scholar]
  17. Nakamura Y., Tanabe K., Egawa K. Structure of major surface determinants and DNA diagnosis of Pneumocystis carinii. J Protozool. 1989 Jan-Feb;36(1):58S–60S. doi: 10.1111/j.1550-7408.1989.tb02699.x. [DOI] [PubMed] [Google Scholar]
  18. Ochs D. Protein contaminants of sodium dodecyl sulfate-polyacrylamide gels. Anal Biochem. 1983 Dec;135(2):470–474. doi: 10.1016/0003-2697(83)90714-5. [DOI] [PubMed] [Google Scholar]
  19. Pifer L. L., Hughes W. T., Murphy M. J., Jr Propagation of Pneumocystis carinii in vitro. Pediatr Res. 1977 Apr;11(4):305–316. doi: 10.1203/00006450-197704000-00010. [DOI] [PubMed] [Google Scholar]
  20. Takeuchi M., Takasaki S., Inoue N., Kobata A. Sensitive method for carbohydrate composition analysis of glycoproteins by high-performance liquid chromatography. J Chromatogr. 1987 Jul 29;400:207–213. doi: 10.1016/s0021-9673(01)81613-7. [DOI] [PubMed] [Google Scholar]
  21. Tschudi C., Ullu E. Polygene transcripts are precursors to calmodulin mRNAs in trypanosomes. EMBO J. 1988 Feb;7(2):455–463. doi: 10.1002/j.1460-2075.1988.tb02833.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Walzer P. D., Linke M. J. A comparison of the antigenic characteristics of rat and human Pneumocystis carinii by immunoblotting. J Immunol. 1987 Apr 1;138(7):2257–2265. [PubMed] [Google Scholar]
  23. Yoshikawa H., Morioka H., Yoshida Y. Ultrastructural detection of carbohydrates in the pellicle of Pneumocystis carinii. Parasitol Res. 1988;74(6):537–543. doi: 10.1007/BF00531631. [DOI] [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES