Immunocytochemical Identification of the Major Exospore Protein and three Polar-Tube Proteins of the Microsporidia Paranosema grylli

Dolgikh, Viacheslav V.; Semenov, Petr; Mirоnоv, Alexander A.; Beznoussenko, Galina V.

doi:10.1016/j.protis.2004.12.002

Cited by 6 publications

(10 citation statements)

References 21 publications

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“…This thus suggests the active participation of the TNs in polar filament formation. Thus, three P. grylli polar-tube proteins (PTP A, PTP B and PTP C) (Dolgikh et al, 2005) should be other examples of secretory proteins that pass through the Golgi.…”

Section: The Tubular Network Contain Golgi Markersmentioning

confidence: 99%

“…To determine whether p40 and the PTP selected as cargo proteins (PTP A) (Dolgikh et al, 2005) undergo posttranslational modifications in the Golgi, we examined whether these two proteins were glycosylated, because glycosylation of transported proteins is one of the main functions of the Golgi. However, this task was not trivial.…”

Section: The Tubular Network Contain Golgi Markersmentioning

confidence: 99%

“…To test this hypothesis, we examined the immunolocalization of p40 and the three PTPs using purified pAbs against these proteins (Dolgikh et al, 2005). In meronts, p40 was only present over the microsporidia ER cisternae (Fig.…”

Section: The Tubular Network Contain Golgi Markersmentioning

confidence: 99%

“…Abs against P. grylli structural proteins of spore wall and polar tube (p40 and PTP A, see below) were generated and purified as described previously (Dolgikh et al, 2005).…”

Section: Antibodiesmentioning

confidence: 99%

“…Human transferrin was incubated under the same conditions as a positive control. To recognize ␣-mannose and ␤-mannose, p40 was selectively extracted from intact spores by their incubation in alkaline-saline solution (10 mM KOH, 170 mM KCl) (Dolgikh et al, 2005), and neutralization with 0.05 volumes of 1.0 M sodium phosphate (pH 6.0) was followed by the incubation either overnight at room temperature or at 37°C for 3 hours in the presence 0.1 U ␣-mannosidase or ␤-mannosidase. The control and enzyme-treated samples were incubated for 1 hour at room temperature in the presence of GNA-lectin crosslinked with agarose beads.…”

Section: Analysis Of Protein Glycosylationmentioning

confidence: 99%

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Analogs of the Golgi complex in microsporidia: structure and avesicular mechanisms of function

Beznoussenko

Dolgikh

Seliverstova

et al. 2007

Journal of Cell Science

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Microsporidia are obligatory intracellular parasites, most species of which live in the host cell cytosol. They synthesize and then transport secretory proteins from the endoplasmic reticulum to the plasma membrane for formation of the spore wall and the polar tube for cell invasion. However, microsporidia do not have a typical Golgi complex. Here, using quick-freezing cryosubstitution and chemical fixation, we demonstrate that the Golgi analogs of the microsporidia Paranosema (Antonospora) grylli and Paranosema locustae appear as 300-nm networks of thin (25- to 40-nm diameter), branching or varicose tubules that display histochemical features of a Golgi, but that do not have vesicles. Vesicles are not formed even if membrane fusion is inhibited. These tubular networks are connected to the endoplasmic reticulum, the plasma membrane and the forming polar tube, and are positive for Sec13, γCOP and analogs of giantin and GM130. The spore-wall and polar-tube proteins are transported from the endoplasmic reticulum to the target membranes through these tubular networks, within which they undergo concentration and glycosylation. We suggest that the intracellular transport of secreted proteins in microsporidia occurs by a progression mechanism that does not involve the participation of vesicles generated by coat proteins I and II.

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Section: The Tubular Network Contain Golgi Markersmentioning

confidence: 99%

Section: The Tubular Network Contain Golgi Markersmentioning

confidence: 99%

Section: The Tubular Network Contain Golgi Markersmentioning

confidence: 99%

“…Abs against P. grylli structural proteins of spore wall and polar tube (p40 and PTP A, see below) were generated and purified as described previously (Dolgikh et al, 2005).…”

Section: Antibodiesmentioning

confidence: 99%

Section: Analysis Of Protein Glycosylationmentioning

confidence: 99%

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Analogs of the Golgi complex in microsporidia: structure and avesicular mechanisms of function

Beznoussenko

Dolgikh

Seliverstova

et al. 2007

Journal of Cell Science

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A proteomic‐based approach for the characterization of some major structural proteins involved in host–parasite relationships from the silkworm parasite Nosema bombycis (Microsporidia)

Wang

Chambon²,

et al. 2007

Proteomics

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Nosema bombycis is the causative agent of the silkworm Bombyx mori pebrine disease which inflicts severe worldwide economical losses in sericulture. Little is known about host-parasite interactions at the molecular level for this spore-forming obligate intracellular parasite which belongs to the fungi-related Microsporidia phylum. Major microsporidian structural proteins from the spore wall (SW) and the polar tube (PT) are known to be involved in host invasion. We developed a proteomic-based approach to identify few N. bombycis proteins belonging to these cell structures. Protein extraction protocols were optimized and four N. bombycis spore protein extracts were compared by SDS-PAGE and 2-DE to establish complementary proteomic profiles. Three proteins were shown to be located at the parasite SW. Moreover, 17 polyclonal antibodies were raised against major N. bombycis proteins from all extracts, and three spots were shown to correspond to polar tube proteins (PTPs) by immunofluorescent assay and transmission electron microscopy immunocytochemistry on cryosections. Specific patterns for each PTP were obtained by MALDI-TOF-MS and MS/MS. Peptide sequence tags were deduced by de novo sequencing using Peaks Online and DeNovoX, then evaluated by MASCOT and SEQUEST searches. Identification parameters were higher than false-positive hits, strengthening our strategy that could be enlarged to a nongenomic context.

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The Function and Structure of the Microsporidia Polar Tube

Han

Takvorian

Weiss

2022

Experientia Supplementum

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Immunocytochemical Identification of the Major Exospore Protein and three Polar-Tube Proteins of the Microsporidia Paranosema grylli

Cited by 6 publications

References 21 publications

Analogs of the Golgi complex in microsporidia: structure and avesicular mechanisms of function

Analogs of the Golgi complex in microsporidia: structure and avesicular mechanisms of function

A proteomic‐based approach for the characterization of some major structural proteins involved in host–parasite relationships from the silkworm parasite Nosema bombycis (Microsporidia)

The Function and Structure of the Microsporidia Polar Tube

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