The inner membrane complex (IMC) of apicomplexan parasites contains a network intermediate filament-like proteins. 14 alveolin domain-containing IMC proteins in Toxoplasma gondii fall into different groups defined by their distinct spatiotemporal dynamics during the internal budding process tachyzoites. Here, we analyzed representatives protein across all stages life cycle and Sarcocystis neurona asexual development. We found that asexually dividing stages, IMC7 is present exclusively...

Significance We describe here the discovery that assemblies of O -fucosylated proteins localize to nuclear membrane Toxoplasma gondii , particularly in proximity pore complexes (NPCs). -fucose is added Ser and Thr residues found some Phe-Gly (FG) domain-containing characterize NPC channel as well Ser-rich sequences many predicted have roles transcription, mRNA processing, cell signaling. -fucosylation nucleocytosolic has not been described previously any eukaryote appears be unique T ....

In many metazoan species, an unusual type of protein glycosylation, called C-mannosylation, occurs on adhesive thrombospondin 1 repeats (TSRs) and I cytokine receptors. This modification has been shown to be catalyzed by the Caenorhabditis elegans DPY-19 orthologues encoding gene were found in genome apicomplexan parasites. Lately, micronemal adhesin thrombospondin-related anonymous (TRAP) was C-hexosylated Plasmodium falciparum sporozoites. Here, we demonstrate that also MIC2 secreted...

The enzymes phosphomannomutase (PMM), phospho-N-acetylglucosamine mutase (PAGM) and phosphoglucomutase (PGM) reversibly catalyse the transfer of phosphate between C6 C1 hydroxyl groups mannose, N-acetylglucosamine glucose respectively. Although genes for a candidate PMM PAGM have been found in Trypanosoma brucei genome, there is, surprisingly, no gene PGM. TbPMM TbPAGM were cloned expressed Escherichia coli enzyme was crystallized its structure solved at 1.85 Å resolution. Antibodies to...

Interconversion of UDP-glucose (UDP-Glc) and UDP-galactose (UDP-Gal) by the UDP-Glc 4´-epimerase intimately connects biosynthesis these two nucleotide sugars. Their de novo involves transformation glucose-6-phosphate into glucose-1-phosphate phosphoglucomutase subsequent activation specific pyrophosphorylase (UGP). Besides UGP, Leishmania parasites express an uncommon UDP-sugar (USP) able to activate both galactose-1-phosphate in vitro. Targeted gene deletion UGP alone was previously shown...

To survive in its sand fly vector, the trypanosomatid protozoan parasite Leishmania first attaches to midgut avoid excretion, but eventually it must detach for transmission by next bite. In major strain Friedlin, this is controlled modifications of stage-specific adhesin lipophosphoglycan (LPG). During differentiation infective metacyclics, d-arabinopyranose (d-Arap) caps LPG side-chain galactose residues, blocking interaction with lectin PpGalec, thereby leading detachment and transmission....

Apicomplexa form a phylum of obligate parasitic protozoa great clinical and veterinary importance. These parasites synthesize glycoconjugates for their survival infectivity, but the enzymatic steps required to generate glycosylation precursors are not completely characterized. In particular, glucosamine-phosphate N-acetyltransferase (GNA1) activity, needed produce essential UDP-N-acetylglucosamine (UDP-GlcNAc) donor, has been identified in any Apicomplexa. We scanned genomes Plasmodium...

Abstract Glycosylation is an important posttranslational protein modification in all eukaryotes. Besides glycosylphosphatidylinositol (GPI) anchors and N -glycosylation, O -fucosylation has been recently reported key sporozoite proteins of the malaria parasite. Previous analyses showed presence GDP-fucose (GDP-Fuc), precursor for fucosylation reactions, blood stages Plasmodium falciparum . The GDP-Fuc de novo pathway, which requires action GDP-mannose 4,6-dehydratase (GMD) GDP-L-fucose...

Leishmaniases are a set of tropical and sub-tropical diseases caused by protozoan parasites the genus Leishmania whose severity ranges from self-healing cutaneous lesions to fatal visceral infections. synthesise wide array cell surface secreted glycoconjugates that play important roles in infection. These particularly abundant promastigote form known be essential for establishment infection insect midgut effective transmission mammalian host. Since they rich galactose, their biosynthesis...

Once considered unusual, nucleocytoplasmic glycosylation is now recognized as a conserved feature of eukaryotes. While in animals, O-GlcNAc transferase (OGT) modifies thousands intracellular proteins, the human pathogen Toxoplasma gondii transfers different sugar, fucose, to proteins involved transcription, mRNA processing, and signaling. Knockout experiments showed that TgSPY, an ortholog plant SPINDLY paralog host OGT, required for nuclear O-fucosylation. Here we verify TgSPY...

Fucose is a common component of eukaryotic cell-surface glycoconjugates, generally added by Golgi-resident fucosyltransferases. Whereas fucosylated glycoconjugates are rare in kinetoplastids, the biosynthesis nucleotide sugar GDP-Fuc has been shown to be essential Trypanosoma brucei. Here we show that single identifiable T. brucei fucosyltransferase (TbFUT1) GDP-Fuc: β-D-galactose α-1,2-fucosyltransferase with an apparent preference for Galβ1,3GlcNAcβ1-O-R acceptor motif. Conditional null...

ABSTRACT Enteric protozoan parasites, which are spread by the fecal-oral route, important causes of diarrhea ( Giardia duodenalis ) and amebic dysentery Entamoeba histolytica ). Cyst walls have a single layer composed fibrils β-1,3-linked GalNAc β-1,4-linked GlcNAc (chitin), respectively. The goal here was to determine whether hand sanitizers that contain ethanol or isopropanol as active microbicide might reduce transmission these parasites. We found treatment with alcohols without drying in...

O-GlcNAcylation is a prominent modification of nuclear and cytoplasmic proteins in animals plants mediated by single O-GlcNAc transferase (OGT). Spindly (Spy), paralog OGT first discovered higher plants, has an ortholog the apicomplexan parasite Toxoplasma gondii, both enzymes are now recognized as O-fucosyltransferases (OFTs). Here we investigate evolution spy-like genes experimentally confirm OFT activity social amoeba Dictyostelium-a protist that more related to fungi metazoa....

Abstract Toxoplasma gondii is an intracellular parasite that causes disseminated infections which can lead to neurological damage in fetuses and immunocompromised individuals. Microneme protein 2 (MIC2) , a member of the thrombospondin-related anonymous (TRAP) family, secreted important for motility, host cell attachment, invasion, egress. MIC2 contains six thrombospondin type I repeats (TSRs) are modified by C-mannose O-fucose Plasmodium spp. mammals. Here we used mass spectrometry show...

ABSTRACT Fucose is a common component of eukaryotic cell-surface glycoconjugates, generally added by Golgi-resident fucosyltransferases. Whereas fucosylated glycoconjugates are rare in kinetoplastids, the biosynthesis nucleotide sugar GDP-Fuc has been shown to be essential Trypanosoma brucei. Here we show that single identifiable T. brucei fucosyltransferase (TbFUT1) GDP-Fuc: β-D-galactose α-1,2-fucosyltransferase with an apparent preference for Galβ1,3GlcNAcβ1-O-R acceptor motif....