Glycoinositolphospholipids, Free and as Anchors of Proteins, in Trypanosoma cruzi

Lederkremer, Rosa M. de; Bertello, Laura E.

doi:10.2174/1381612013397519

Cited by 35 publications

(27 citation statements)

References 58 publications

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“…Both GIPLs differ in the lipid moiety, whereas 1-O-hexadecyl-2-O-palmitoyl glycerol was identified in GIPL A, ceramides, mainly N-palmitoyl and N-lignoceryl sphinganines, were found in GIPL B. The ceramides are the same as those found in the LPPG [29]. It is known that antibodies against LPPG recognize the Galf residues in these molecules [17].…”

Section: Discussionmentioning

confidence: 94%

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Phospholipid and glycolipid composition of acidocalcisomes of Trypanosoma cruzi

Salto

Kuhlenschmidt

et al. 2008

Molecular and Biochemical Parasitology

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Highly purified acidocalcisomes from Trypanosoma cruzi epimastigotes were obtained by differential centrifugation and iodixanol gradient ultracentrifugation. Lipid analysis of acidocalcisomes revealed the presence of low amounts of 3β-hydroxysterols and predominance of phospholipids. Alkylacyl phosphatidylinositol (16:0/18:2), diacyl phosphatidylinositol (18:0/18:2), diacyl phosphatidylcholine (16:0/18:2; 16:1/18:2; 16:2/18:2, 18:1/18:2, and 18:2/18:2), and diacyl phosphatidylethanolamine (16:0/18:2 and 16:1/18:2) were the only phospholipids characterized by electrospray ionization-mass spectrometry (ESI-MS). Incubation of epimastigotes with [ 3 H]-mannose and isolation of acidocalcisomes allowed the detection of a glycoinositolphospholipid (GIPL) in these organelles. The sugar content of the acidocalcisomal GIPL was similar to that of the GIPL present in a microsomal fraction but the amount of galactofuranose and inositol with respect to the other monosaccharides was lower, suggesting a different chemical structure. Taken together, these results indicate that acidocalcisomes of T. cruzi have a distinct lipid and carbohydrate composition.

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Section: Discussionmentioning

confidence: 94%

“…The purity of the acidocalcisome fraction (fraction 16) was evaluated with plasma membrane (adenylyl cyclase; [28]; P-type H + -ATPase; [22], and LPPG; [29]), glycosome (hexokinase; [30]), and acidocalcisome (V-H + -PPase, [12]) markers. Fig.…”

Section: Isolation Of Microsomal and Acidocalcisome Fractionsmentioning

confidence: 99%

Phospholipid and glycolipid composition of acidocalcisomes of Trypanosoma cruzi

Salto

Kuhlenschmidt

et al. 2008

Molecular and Biochemical Parasitology

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“…The non-protein-linked GPI structures include the so-called glycoinositolphospholipids, or GIPLs, that are most abundant in the leishmania and T. cruzi species (58,64,90) but are also found in procyclic-form T. brucei (60,73,122) and bloodstream form T. brucei (39,63). Non-protein-linked GPIs also include the more exotic leishmania-specific lipophosphoglycan (LPG) structures (38,47,64,65).…”

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confidence: 99%

Sugar Nucleotide Pools of Trypanosoma brucei , Trypanosoma cruzi , and Leishmania major

2007

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The cell surface glycoconjugates of trypanosomatid parasites are intimately involved in parasite survival, infectivity, and virulence in their insect vectors and mammalian hosts. Although there is a considerable body of work describing their structure, biosynthesis, and function, little is known about the sugar nucleotide pools that fuel their biosynthesis. In order to identify and quantify parasite sugar nucleotides, we developed an analytical method based on liquid chromatography-electrospray ionization-tandem mass spectrometry using multiple reaction monitoring. This method was applied to the bloodstream and procyclic forms of Trypanosoma brucei, the epimastigote form of T. cruzi, and the promastigote form of Leishmania major. The trypanosomatids are protozoan parasites that impose a major health burden on many countries in the developing world, causing a wide range of diseases over three continents. Like most eukaryotes, the cell surfaces and endosomal/lysosomal systems of these organisms are rich in glycoconjugates, some of which play essential roles in their survival, infectivity, or virulence. The glycoconjugate repertoires of the three trypanosomatids studied here, Trypanosoma brucei, T. cruzi, and Leishmania major, are fundamentally different, reflecting their disparate life cycles, modes of infection, and disease pathologies (see references 5,27,28,38,47,65, and 69 and references therein). The monosaccharides that make up these glycoconjugates vary between the three trypanosomatid species. For example, all three contain D-mannose (Man), D-Nacetylglucosamine (GlcNAc), D-glucosamine (GlcN), D-glucose (Glc), and D-galactopyranose (Galp), while only T. cruzi and L. major contain D-galactofuranose (Galf), only T. cruzi contains D-xylose (Xyl), L-rhamnopyranose (Rha), and L-fucose (Fuc), and only L. major contains D-arabinopyranose (Ara) ( Table 1). However, a unifying theme of their glycobiology is an abundance of cell surface glycosylphosphatidylinositol (GPI)-anchored glycoproteins and/or non-proteinlinked GPI structures.The protein-linked GPI glycans of the leishmania are the simplest in structure, consisting of the conserved Man␣1-2Man␣1-6Man␣1-4GlcN core. Those of T. cruzi are principally Man␣1-2Man␣1-2Man␣1-6Man␣1-4GlcN, and those of T. brucei are the most complex, with ␣-D-Galp and ␤-D-Galp side chains in the bloodstream form and sialylated poly-N-acetyllactosamine (poly-LacNAc) side chains in the procyclic form (27).The non-protein-linked GPI structures include the so-called glycoinositolphospholipids, or GIPLs, that are most abundant in the leishmania and T. cruzi species (58, 64, 90) but are also found in procyclic-form T. brucei (60,73,122) and bloodstream form T. brucei (39,63). Non-protein-linked GPIs also include the more exotic leishmania-specific lipophosphoglycan (LPG) structures (38,47,64,65). The leishmania LPGs contain characteristic phosphosaccharide repeats of Galp␤1-4Man␣1-P, which in L. major can have ␤-D-Galp and ␣-D-Arap side chains (66,67). These repeats are also found in the secr...

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“…Triplicate samples of NETNES (from 2.8 ϫ 10 9 cell eq) were mixed with 20 pmol of scyllo-inositol internal standard and subjected to GC-MS monosaccharide analysis (25). Triplicate samples of NETNES (from 2.8 ϫ 10 9 cell eq) were mixed with 20 pmol of myo- [1,2,3,4,5, H]inositol internal standard and subjected to GC-MS myo-inositol quantification (25). GC-MS was performed on a Hewlett-Packard 6890-5973 system using an Agilent Technologies HP5 column (30 m ϫ 0.25 mm).…”

Section: Methodsmentioning

confidence: 99%

Structural Characterization of NETNES, a Novel Glycoconjugate in Trypanosoma cruzi Epimastigotes

MacRae

Acosta-Serrano

Morrice

et al. 2005

Journal of Biological Chemistry

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The unicellular stercorarian protozoan parasite Trypanosoma cruzi is the etiological agent of Chagas' disease. The epimastigote form of the parasite is covered in a dense coat of glycoinositol phospholipids and short glycosylphosphatidylinositol (GPI)-anchored mucinlike molecules. Here, we describe the purification and structural characterization of NETNES, a relatively minor but unusually complex glycoprotein that coexists with these major surface components. The mature glycoprotein is only 13 amino acids in length, with the sequence AQENETNESGSID, and exists in two forms with either four or five post-translational modifications. These are either one or two asparagine-linked oligomannose glycans, two linear ␣-mannose glycans linked to serine residues via phosphodiester linkages, and a GPI membrane anchor attached to the C-terminal aspartic acid residue. The variety and density of post-translational modifications on an unusually small peptide core make NETNES a unique type of glycoprotein. The N-glycans are predominantly Man␣1-6(Man␣1-3) Man␣1-6(Man␣1-3)Man␤1-4GlcNAc␤1-4GlcNAc␤1-Asn; the phosphate-linked glycans are a mixture of (Man␣1-2) 0 -3 Man1-P-Ser; and the GPI anchor has the structure Man␣1-2(ethanolamine phosphate)Man␣1-2Man␣1-6Man␣1-4(2-aminoethylphosphonate-6)GlcN␣1-6-myoinositol-1-P-3(sn-1-O-(C 16:0 )alkyl-2-O-(C 16:0 )acylglycerol). Four putative NETNES genes were found in the T. cruzi genome data base. These genes are predicted to encode 65-amino acid proteins with cleavable 26-amino acid Nterminal signal peptides and 26-amino acid C-terminal GPI addition signal peptides.

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Glycoinositolphospholipids, Free and as Anchors of Proteins, in Trypanosoma cruzi

Cited by 35 publications

References 58 publications

Phospholipid and glycolipid composition of acidocalcisomes of Trypanosoma cruzi

Phospholipid and glycolipid composition of acidocalcisomes of Trypanosoma cruzi

Sugar Nucleotide Pools of Trypanosoma brucei , Trypanosoma cruzi , and Leishmania major

Structural Characterization of NETNES, a Novel Glycoconjugate in Trypanosoma cruzi Epimastigotes

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