Cell wall antigens of Pneumocystis carinii trophozoites and cysts

Purification and carbohydrate analysis of these glycoproteins

Jamie Ann De Stefano, John D. Myers, David Du Pont, Jillana M. Foy, Sue A. Theus, Peter D. Walzer

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

We have purified and biochemically analyzed individual cell wall glycoproteins of Pneumocystis carinii. Our results show that corresponding core glycoproteins constitute the cell wall antigens in both trophozoites and cysts, and glycosylation of these glycoproteins does not appear to be significantly altered during development. Cysts and trophozoites in rat-derived organism preparations were separated from each other by counterflow centrifugal elutriation, then treated with Zymolyase to obtain the cell wall fractions. Gel electrophoresis patterns of these fractions from both life-cycle stages were qualitatively similar. Ten major antigenic glycoproteins in these fractions were purified by preparative continuous elution gel electrophoresis. All ten glycoproteins from cysts and trophozoites contained mannose, glucose, galactose, and N-acetylglucosamine, and some contained traces of fucose. The glycoproteins of cysts had more mannose than their trophozoite counterparts. The trophozoite glycoproteins differed from those of the cyst by the presence of xylose. To examine the species-specificity of glycoprotein glycosylation, preparations of human-derived P. carinii (comprised of mixed life-cycle stages) were also examined and found to contain the same sugars as those found in rat-derived organisms. Most of the purified rat-derived glycoproteins bound Concanavalin A, which was abolished by treatment with N-glycanase. This suggested that the majority of the oligosaccharides were N-linked to the proteins, but attempts to identify carbohydrate linkage sites by amino acid sequencing were hampered by apparent modifications of residues. The peptides derived by cyanogen bromide cleavage revealed distinct size patterns for each glycoprotein, suggesting that they were distinct proteins. Most of the glycoproteins reacted with monoclonal antibodies which recognize a highly conserved epitope on rat P. carinii. Four of the individually purified glycoprotein preparations elicited in vitro cellular immune responses, implicating their involvement in the recognition of P. carinii by host T cells. The identification and characterization of P. carinii cell wall proteins will be helpful in analyzing the relationship of the organism to its mammalian host.

Original languageEnglish (US)
Pages (from-to)334-343
Number of pages10
JournalJournal of Eukaryotic Microbiology
Volume45
Issue number3
StatePublished - May 1 1998

Fingerprint

Pneumocystis carinii
Trophozoites
Cell Wall
Cysts
Glycoproteins
Carbohydrates
Antigens
Mannose
Life Cycle Stages
Glycosylation
Electrophoresis
Carbohydrate Conformation
Gels
Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase
Species Specificity
Cyanogen Bromide
Proteins
Fucose
Acetylglucosamine
Xylose

Keywords

  • Biochemical analysis
  • Developmental stages
  • Opportunistic pathogen
  • Structure

ASJC Scopus subject areas

  • Microbiology

Cite this

Cell wall antigens of Pneumocystis carinii trophozoites and cysts : Purification and carbohydrate analysis of these glycoproteins. / De Stefano, Jamie Ann; Myers, John D.; Du Pont, David; Foy, Jillana M.; Theus, Sue A.; Walzer, Peter D.

In: Journal of Eukaryotic Microbiology, Vol. 45, No. 3, 01.05.1998, p. 334-343.

Research output: Contribution to journalArticle

De Stefano, Jamie Ann ; Myers, John D. ; Du Pont, David ; Foy, Jillana M. ; Theus, Sue A. ; Walzer, Peter D. / Cell wall antigens of Pneumocystis carinii trophozoites and cysts : Purification and carbohydrate analysis of these glycoproteins. In: Journal of Eukaryotic Microbiology. 1998 ; Vol. 45, No. 3. pp. 334-343.
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