The molecular characterization of the major polar tube protein gene from Encephalitozoon hellem, a microsporidian parasite of humans

Elaine M. Keohane, George A. Orr, Hong Shan Zhang, Peter M. Takvorian, Ann Cali, Herbert B. Tanowitz, Murray Wittner, Louis M. Weiss

Research output: Contribution to journalArticle

56 Citations (Scopus)

Abstract

The microsporidia are obligate intracellular protozoan parasites of increasing importance as human pathogens, which are characterized by a small resistant spore with a single polar filament that coils around the sporoplasm. When stimulated, the polar filament rapidly everts out of the spore to form a hollow polar tube through which the sporoplasm passes, thus serving as a unique mechanism of transmission. A genomic library of the human microsporidium Encephalitozoon hellem was screened using a polyclonal rabbit antibody (anti-PTP Eh55) produced to the major HPLC purified polar tube protein (PTP) of E. hellem. This antibody localized to intrasporal polar filaments and extrasporal polar tubes of E. hellem by immunogold electron microscopy confirming the polar tube specificity of the antibody. A total of 14 anti-PTP Eh55 reactive genomic clones were identified and purified. A PTP gene was identified consisting of 1362 bp coding for 453 amino acids. The N-terminus of the translated protein consists of a putative N-terminal signal sequence of 22 amino acids, which when cleaved results in a mature protein of 431 amino acids with a predicted molecular mass of 43 kDa. The protein has a high proline content (14.6%) and contains a central domain of six alternating tandem repeats of 20 amino acids. After ligation of the gene into a glutathione S-transferase (GST) expression vector, a fusion protein was produced that reacted by immunoblotting with the polar tube specific anti-PTP Eh55. The gene was present as a single copy in the genome and there was no homology with other known genes. As the polar tube is a critical structure for the transmission of this organism to a new host cell, further study of PTPs may lead to the development of new therapeutic strategies and diagnostic tests. Copyright (C) 1998 Elsevier Science B.V.

Original languageEnglish (US)
Pages (from-to)227-236
Number of pages10
JournalMolecular and Biochemical Parasitology
Volume94
Issue number2
DOIs
StatePublished - Aug 1 1998

Fingerprint

Parasites
Proteins
Amino Acids
Spores
Unclassified Microsporidia
Encephalitozoon
Microsporidia
Genes
Encephalitozoon hellem PTP protein
Tandem Repeat Sequences
Antibody Specificity
Genomic Library
Protein Sorting Signals
Glutathione Transferase
Immunoblotting
Routine Diagnostic Tests
Proline
Ligation
Anti-Idiotypic Antibodies
Electron Microscopy

Keywords

  • Encephalitozoon
  • Microsporidia
  • Polar filament
  • Polar tube
  • Polar tube protein
  • Polar tube protein gene

ASJC Scopus subject areas

  • Molecular Biology
  • Parasitology

Cite this

The molecular characterization of the major polar tube protein gene from Encephalitozoon hellem, a microsporidian parasite of humans. / Keohane, Elaine M.; Orr, George A.; Zhang, Hong Shan; Takvorian, Peter M.; Cali, Ann; Tanowitz, Herbert B.; Wittner, Murray; Weiss, Louis M.

In: Molecular and Biochemical Parasitology, Vol. 94, No. 2, 01.08.1998, p. 227-236.

Research output: Contribution to journalArticle

Keohane, Elaine M. ; Orr, George A. ; Zhang, Hong Shan ; Takvorian, Peter M. ; Cali, Ann ; Tanowitz, Herbert B. ; Wittner, Murray ; Weiss, Louis M. / The molecular characterization of the major polar tube protein gene from Encephalitozoon hellem, a microsporidian parasite of humans. In: Molecular and Biochemical Parasitology. 1998 ; Vol. 94, No. 2. pp. 227-236.
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