Identification of a microsporidian polar tube protein reactive monoclonal antibody

Elaine M. Keohane, Peter M. Takvorian, Ann Cali, Herbert B. Tanowitz, Murray Wittner, Louis M. Weiss

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

The microsporidia are characterized by spores containing a single polar tube that coils around the sporoplasm. When triggered by appropriate stimuli, the polar tube rapidly discharges out of the spore forming a hollow tube. The sporoplasm passes out of the spore through this tube serving as a unique vehicle of infection. Due to the unusual functional and solubility properties of the polar tube, the proteins comprising it are likely to be members of a protein family with a highly conserved amino acid composition among the various microsporidia. Polar tube proteins were separated from the majority of other proteins in glass bead disrupted spores of Glugea americanus using sequential 1% sodium dodecyl sulfate (SDS) and 9M urea extractions. The resultant spore pellet demonstrated broken, empty spore coats and numerous polar tubes in straight and twisted formations by negative stain transmission electron microscopy. After subsequent incubation of the pellet with 2% dithiothreitol (DTT), empty spore coats were still observed but the polar tubes were no longer present in the pellet. The DTT supernatant demonstrated four major protein bands by SDS-PAGE: 23, 27, 34 and 43 kDa. Monoclonal antibodies were produced to these proteins using Hunter's Titermax adjuvant. Mab 3C8.23.1 which cross-reacted with a 43-kDa antigen by immunoblot analyis, demonstrated strong reactivity with the polar tube of G. americanus spores by immunogold electron microscopy. This antibody will be useful in further characterization of polar tube proteins and may lead to novel diagnostic and therapeutic reagents.

Original languageEnglish (US)
Pages (from-to)26-31
Number of pages6
JournalJournal of Eukaryotic Microbiology
Volume43
Issue number1
StatePublished - Jan 1996

Fingerprint

Microsporidia
Spores
monoclonal antibodies
spores
Monoclonal Antibodies
Proteins
proteins
pellets
dithiothreitol
Dithiothreitol
sodium dodecyl sulfate
Glugea
Transmission Electron Microscopy
amino acid composition
Sodium Dodecyl Sulfate
Solubility
adjuvants
Glass
polyacrylamide gel electrophoresis
transmission electron microscopy

Keywords

  • Glugea americanus
  • Immunogold electron microscopy
  • Microspora
  • Microsporidia
  • Polar filament

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Agricultural and Biological Sciences (miscellaneous)
  • Applied Microbiology and Biotechnology
  • Microbiology

Cite this

Keohane, E. M., Takvorian, P. M., Cali, A., Tanowitz, H. B., Wittner, M., & Weiss, L. M. (1996). Identification of a microsporidian polar tube protein reactive monoclonal antibody. Journal of Eukaryotic Microbiology, 43(1), 26-31.

Identification of a microsporidian polar tube protein reactive monoclonal antibody. / Keohane, Elaine M.; Takvorian, Peter M.; Cali, Ann; Tanowitz, Herbert B.; Wittner, Murray; Weiss, Louis M.

In: Journal of Eukaryotic Microbiology, Vol. 43, No. 1, 01.1996, p. 26-31.

Research output: Contribution to journalArticle

Keohane, EM, Takvorian, PM, Cali, A, Tanowitz, HB, Wittner, M & Weiss, LM 1996, 'Identification of a microsporidian polar tube protein reactive monoclonal antibody', Journal of Eukaryotic Microbiology, vol. 43, no. 1, pp. 26-31.
Keohane, Elaine M. ; Takvorian, Peter M. ; Cali, Ann ; Tanowitz, Herbert B. ; Wittner, Murray ; Weiss, Louis M. / Identification of a microsporidian polar tube protein reactive monoclonal antibody. In: Journal of Eukaryotic Microbiology. 1996 ; Vol. 43, No. 1. pp. 26-31.
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