Labeling and initial characterization of polar lipids in cultures ofPlasmodium falciparum

Dieckmann-Schuppert, Angela; Bender, Stephanie; Holder, Anthony A.; Haldar, Kasturi; Schwarz, Ralph Τ.

doi:10.1007/bf00931698

Cited by 11 publications

(7 citation statements)

References 44 publications

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“…1 and Table 1), although redundant activity from PF3D7_ 0517500 might be expected. The lack of a candidate for a gene encoding for UDP-glucose 4-epimerase (EC 5.1.3.2) that could present a UDP-GlcNAc 4-epimerase activity (EC 5.1.3.7) to convert UDP-GlcNAc into UDP-GalNAc indicates that the parasite probably does not produce UDP-N-acetyl galactosamine, in agreement with the inability of the parasite to synthesize GalNAc and the absence of mucin-type O-glycosylation in P. falciparum (63,65).…”

Section: Discussionsupporting

confidence: 53%

“…2), along with the labeling of GPI-anchored proteins using tritiated glucosamine (Fig. 3C) (23,63), strongly suggests that the UDP-GlcNAc metabolic route (25) (Fig. 1) is active in P. falciparum.…”

Section: Discussionmentioning

confidence: 94%

“…Interestingly, searches with functionally characterized UDP-sugar pyrophosphorylase orthologues from plants and L. major identify a match, PF3D7_0517500, with more than 30% similarity and an e value of Ͻ10 Ϫ55 . However, although galactose competes for P. falciparum PfHT1 hexose permease (45) and its incorporation into the parasite galactolipids has been reported (62,63), there is not a clear galactokinase candidate in the P. falciparum genome, and tritiated galactose is not significantly incorporated into the parasite proteins (23) or into the ␣-galactose moieties of digalactosyl diglycerides (64).…”

Section: Discussionmentioning

confidence: 99%

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Biosynthesis of GDP-fucose and Other Sugar Nucleotides in the Blood Stages of Plasmodium falciparum

Sanz

Bandini

Ospina

et al. 2013

Journal of Biological Chemistry

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Background: GDP-fucose and other sugar nucleotide biosynthetic pathways are conserved in the P. falciparum genome. Results: These pathways are active in the intraerythrocytic life cycle of the parasite. Conclusion:The parasite biosynthesizes GDP-fucose and other sugar nucleotides not related to the glycosylphosphatidylinositol structures Significance: Their presence strongly suggests that they are involved in the biosynthesis of glycans not yet characterized.

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Section: Discussionsupporting

confidence: 53%

Section: Discussionmentioning

confidence: 94%

Section: Discussionmentioning

confidence: 99%

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Biosynthesis of GDP-fucose and Other Sugar Nucleotides in the Blood Stages of Plasmodium falciparum

Sanz

Bandini

Ospina

et al. 2013

Journal of Biological Chemistry

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“…Lipids were removed from 50 pl lysate by three subsequent extractions with 3 ml ice-cold hexanelisopropanol (3:2, by vol. ; Dieckmann-Schuppert et al, 1992b). The residue was then exhaustively treated with protein N-glycanase, which releases 7-10% of the GlcN label, the nature of which is still unknown (Dieckmann-Schuppert et al, 1992a).…”

Section: Sample Preparationmentioning

confidence: 99%

Studies on O‐glycans of Plasmodium‐falciparum‐infected human erythrocytes Evidence for O‐GlcNAc and O‐GlcNAc‐transferase in malaria parasites

Dieckmann-Schuppert

Bause

Schwarz

1993

European Journal of Biochemistry

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) -EJB 930465/4 0-Glycosylation is the major form of protein glycosylation in human erythrocytes infected with the asexual intraerythrocytic stage of the malaria parasite, Plasmodium falciparum. This study compares aspects of 0-glycosylation in l? falciparum-infected and uninfected erythrocytes.Non-labeled and metabolically glucosamine-labeled 0-glycans were obtained from the protein fraction of infected or uninfected erythrocytes by p elimination. Additional label was introduced by reduction with sodium borohydride, or by the attachment of radioactive Gal to peripheral GlcNAc using galactosyltransferase. 2 -4-times more labeled 0-glycans were obtained from infected erythrocytes compared to the same number of uninfected ones, consistent with additional biosynthesis by the parasite. Our analysis of these 0-glycans showed no significant qualitative divergence between the 0-glycans of the infected and those of the uninfected red cell.According to preliminary alditol analyses, the 0-glycans of P. falciparum-infected red cells do not contain GalNAc at their reducing terminus. Moreover, GalNAc was not synthesized by P. falciparum from either Glc, Gal, GlcN or GalN. At least one 0-glycan found in P. faleiparurn-infected erythrocytes contains GlcNAc at its reducing terminus.Gel-filtration results had suggested the presence of 0-GlcNAc on proteins in the infected erythrocyte. Probing with a synthetic pentapeptide, we could show that P. falciparum expresses its own 0-GlcNAc transferase during intraerythrocytic development. Using this peptide, the enzyme was characterized to some degree. The localization and function of 0-GlcNAc in P. falciparum remains to be elucidated.Plasmodium falciparum is the causative agent of human malignant malaria tropica. Despite huge efforts in vaccine and chemotherapy development, this disease still causes the death of several million people each year. A more thorough understanding of the biochemistry and cell biology of this parasite is required in order to develop better chemotherapy and vaccination strategies. One of the neglected areas of malaria biochemistry is the glycobiology of the parasite. Little is known about the biological significance of oligosaccharides in I? falciparum, be they linked to lipids or to proteins.However, post-translational modifications of proteins by the covalent attachment of sugars are often very important for the function, localization and eventual antigenicity of proteins concerned (Rademacher et al., 1988 We have previously shown that glycoproteins of the asexual intraerythrocytic P. fulciparum bear mainly, if not exclusively, 0-glycans, whereas N-glycans seem to be lacking (Dieckmann-Schuppert et al., 1992a). The present study was undertaken in order to investigate the nature of the 0-glycans present in P. faleiparum-infected erythrocytes in general and the occurrence of 0-GlcNAc in particular.Despite the possibility of growing P. falciparum continuously in culture (Trager and Jensen, 1976), the biochemical analysis of this system is hampered by the facts t...

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“…Hydrophobic (glyco)lipids, which could, for example, be dolichol cycle or glycosyl-PtdIns-anchor biosynthetic intermediates, were studied by TLC analysis after organic solvent extraction from metabolically labeled cultures (Dieckmann-Schuppert et al, 1992b). N o difference was seen when the cultures were radiolabeled with tritiated glucosamine, palmitate, myristate or inositol, in the presence or absence of tunicamycin (data not shown).…”

Section: Fig 3 Gel Filtration Analysis On Sephadex G25 Of P Falcipmentioning

confidence: 99%

Apparent lack of N‐glycosylation in the asexual intraerythrocytic stage of Plasmodium falciparum

Dieckmann-Schuppert

Bender

Odenthal-Schnittler

et al. 1992

European Journal of Biochemistry

119

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This study investigates protein glycosylation in the asexual intraerythrocytic stage of the malaria parasite, Plasmodium ,fakiparum, and the presence in the infected erythrocyte of the respective precursors.In in vitro cultures, P. fakiparum can be metabolically labeled with radioactive sugars, and its multiplication can be affected by glycosylation inhibitors, suggesting the capability of the parasite to perform protein-glycosylation reactions. Gel-filtration analysis of sugar-labeled malarial proteins before and after specific cleavage of N-glycans or 0-glycans, respectively, revealed the majority of the protein-bound sugar label to be incorporated into 0-glycans, but only little (7-12% of the glucosamine label) or no N-glycans were found. Analysis of the nucleotide sugar and sugar-phosphate fraction showed that radioactive galactose, glucosamine, fucose and ethanolamine were converted to their activated derivatives required for incorporation into protein. Mannose was mainly recovered as a bisphosphate, whereas the level of radiolabeled GDP-mannose was below the detection limit. The analysis of organic-solvent extracts of sugar-labeled cultures showed no evidence for the formation by the parasite of dolichol cycle intermediates, the dedicated precursors in protein N-glycosylation. Consistently, the amount of UDP-N-acetylglucosamine formed did not seem to be affected by the presence of tunicamycin in the culture. Oligosaccharyl-transferase activity was not detectable in a lysate of P. fakiparum, using exogenous glycosyl donors and acceptors.Our studies show that 0-glycosylation is the major form of protein glycosylation in intrderythrocytic P. julciparum, whereas there is little or no protein N-glycosylation. A part of these studies has been published in abstract form [Dieckmann-Schuppert, A,, Hensel, J. and Schwarz, R. T. (1991) Biol. Chem. Hoppe-Seyler 372,6451.Plasmodium fakiparum is the causative agent of human malignant malaria tropica. Despite huge efforts in vaccine and chemotherapy development, today this disease still causes the death of several million people/year (World Health Organization, 1989). A more thorough understanding of the biochemistry and cell biology of this parasite is required in order to develop better chemotherapy and vaccination strategies. One of the neglected areas of malaria biochemistry is the glycobiology of the parasite. Very little is known to date about the biological significance of oligosaccharides in P. julcipurum, be they linked to lipids or to proteins. Glycolipids may be membrane components and as such be potential antigens, or be involved in the formation of glycoproteins, e. g. dolichol

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Labeling and initial characterization of polar lipids in cultures ofPlasmodium falciparum

Cited by 11 publications

References 44 publications

Biosynthesis of GDP-fucose and Other Sugar Nucleotides in the Blood Stages of Plasmodium falciparum

Biosynthesis of GDP-fucose and Other Sugar Nucleotides in the Blood Stages of Plasmodium falciparum

Studies on O‐glycans of Plasmodium‐falciparum‐infected human erythrocytes Evidence for O‐GlcNAc and O‐GlcNAc‐transferase in malaria parasites

Apparent lack of N‐glycosylation in the asexual intraerythrocytic stage of Plasmodium falciparum

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