Characterization of prenylated protein methyltransferase in Leishmania

Hasne, Marie Pierre; Lawrence, Françoise

doi:10.1042/bj3420513

Cited by 7 publications

(3 citation statements)

References 46 publications

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“…In order to obtain a fraction enriched with the plasma membrane, cell fractionation was performed by differential centrifugation by the method of Hasne and Lawrence (10). Briefly, promastigotes were grown in 1 liter of medium as described above, and the parasites were harvested by centrifugation at 4,000 ϫ g for 5 min at 4°C and then washed three times with cold TBS and resuspended in 100 mM Tris/HCl, pH 7.5, containing 1 mM EDTA and 250 mM sucrose.…”

Section: Methodsmentioning

confidence: 99%

Miltefosine Affects Lipid Metabolism in Leishmania donovani Promastigotes

Rakotomanga

Blanc

Gaudin

et al. 2007

Antimicrob Agents Chemother

150

120

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Miltefosine (hexadecylphosphocholine [ HePC]) is the first orally active antileishmanial drug. Transient HePC treatment of Leishmania donovani promastigotes at 10 M significantly reduced the phosphatidylcholine content and enhanced the phosphatidylethanolamine (PE) content in parasite membranes, suggesting a partial inactivation of PE-N-methyltransferase. Phospholipase D activity did not seem to be affected by HePC. In addition, the enhancement of the lysophosphatidylcholine content could be ascribed to phospholipase A2 activation. Moreover, transient HePC treatment had no effect on the fatty acid alkyl chain length or the fatty acid unsaturation rate. Concerning sterols, we found a strong reduction of the C 24 alkylated sterol content, and the enhancement of the cholesterol content could be the result of the HePC condensation effect with sterols.

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Section: Methodsmentioning

confidence: 99%

Miltefosine Affects Lipid Metabolism in Leishmania donovani Promastigotes

Rakotomanga

Blanc

Gaudin

et al. 2007

Antimicrob Agents Chemother

150

120

View full text Add to dashboard Cite

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“…Cell fractionation by differential centrifugation was performed according to the method of Hasne and Lawrence (8). Briefly, promastigotes were grown in 1 liter of medium as described above, and for HePC-R parasites, drug pressure was stopped in the subculture preceding the experiment to avoid drug contamination.…”

Section: Methodsmentioning

confidence: 99%

Alteration of Fatty Acid and Sterol Metabolism in Miltefosine-Resistant Leishmania donovani Promastigotes and Consequences for Drug-Membrane Interactions

Rakotomanga

Saint-Pierre-Chazalet

Loiseau

2005

Antimicrob Agents Chemother

130

113

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Miltefosine (hexadecylphosphocholine [HePC]) is the first orally active drug approved for the treatment of visceral leishmaniasis. In order to investigate the biochemical modifications occurring in HePC-resistant (HePC-R) Leishmania donovani promastigotes, taking into account the lipid nature of HePC, we investigated their fatty acid and sterol metabolisms. We found that the content of unsaturated phospholipid alkyl chains was lower in HePC-R parasite plasma membranes than in those of the wild type, suggesting a lower fluidity of HePC-R parasite membranes. We also demonstrated that HePC insertion within an external monolayer was more difficult when the proportion of unsaturated phospholipids decreased, rendering the HePC interaction with the external monolayer of HePC-R parasites more difficult. Furthermore, HePC-R parasite membranes displayed a higher content of short alkyl chain fatty acids, suggesting a partial inactivation of the fatty acid elongation enzyme system in HePC-R parasites. Sterol biosynthesis was found to be modified in HePC-R parasites, since the 24-alkylated sterol content was halved in HePC-R parasites; however, this modification was not related to HePC sensitivity. In conclusion, HePC resistance affects three lipid biochemical pathways: fatty acid elongation, the desaturase system responsible for fatty acid alkyl chain unsaturation, and the C-24-alkylation of sterols.Leishmania spp. are the protozoan parasites responsible for leishmaniases, a family of diseases including a variety of clinical manifestations classically labeled as visceral, cutaneous, and mucocutaneous leishmaniases (9). Chemotherapy is the main tool for the control of leishmaniasis; however, the standard treatments, including pentavalent antimonials, amphotericin B (AmB), and paromomycin, are toxic and expensive.

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“…After the action of either PFT or PGGT-I, a prenyl protein-specific protease cleaves the terminal tripeptide from the prenylated protein (24). The final step is methylation of the terminal carboxylic acid by a prenyl protein-specific methyltransferase (25)(26)(27). Both of these subsequent enzymatic steps have been shown to be required for the proper localization of certain mammalian proteins and are currently being investigated as additional chemotherapeutic targets (28,29).…”

Section: Protein Prenylation In Higher Eukaryotic Cellsmentioning

confidence: 99%

Thematic review series: Lipid Posttranslational Modifications. Fighting parasitic disease by blocking protein farnesylation

Eastman

Buckner

Yokoyama

et al. 2006

Journal of Lipid Research

103

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Protein farnesylation is a form of posttranslational modification that occurs in most, if not all, eukaryotic cells. Inhibitors of protein farnesyltransferase (PFTIs) have been developed as anticancer chemotherapeutic agents. Using the knowledge gained from the development of PFTIs for the treatment of cancer, researchers are currently investigating the use of PFTIs for the treatment of eukaryotic pathogens. This "piggy-back" approach not only accelerates the development of a chemotherapeutic agent for protozoan pathogens but is also a means of mitigating the costs associated with de novo drug design. PFTIs have already been shown to be efficacious in the treatment of eukaryotic pathogens in animal models, including both Trypanosoma brucei, the causative agent of African sleeping sickness, and Plasmodium falciparum, one of the causative agents of malaria. Here, current evidence and progress are summarized that support the targeting of protein farnesyltransferase for the treatment of parasitic diseases. Parasitic diseases continue to have a major impact on morbidity and mortality in tropical and subtropical regions. Among these, malaria causes $300 million infections annually, with 1-3 million deaths occurring in Africa (1). The emergence and spread of parasites resistant to existing antimalarial agents is largely responsible for the recent increase in malaria-related mortality. Another reemerging disease is African sleeping sickness (African trypanosomiasis), with an estimated 50,000 deaths in 2002 (1). The increasing burden of these diseases, along with the inadequacies of current drugs for African sleeping sickness in terms of safety, efficacy, and ease of administration, have led investigators to seek new chemotherapeutic agents (2, 3). Among the current drug targets under study are enzymes involved in protein prenylation, or the posttranslational modification of proteins by the covalent modification by isoprenyl lipids, C15 farnesyl and C20 geranylgeranyl (4-7). The isoprenyl lipid modification of proteins has been shown to be critical for various cellular activities in mammals and yeast, including proliferation and apoptosis (8, 9). Growth of the protozoan parasites has been shown to be severely impaired by the inhibition of protein farnesylation compared with mammalian cells, suggesting high potential of the enzyme protein farnesyltransferase (PFT) as an antiparasitic drug target (5, 10-13).The isoprenoid synthesis pathway from mevalonic acid in many eukaryotes, including trypanosomatids (or deoxyxylulose in Apicomplexa, including Plasmodium and Toxoplasma, and plants) is essential for the production of sterols, dolichol, ubiquinone, and other isoprene derivatives in many eukaryotic cells. Indeed, these pathways have been the study of recent efforts to develop other antiparasitic chemotherapeutic agents, especially the targeting of isoprenoid pyrophosphate synthesis by nitrogen-containing bisphosphonates (14-16). Organisms belonging to the group Apicomplexa contain the nonmevalonate pathway of iso...

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Characterization of prenylated protein methyltransferase in Leishmania

Cited by 7 publications

References 46 publications

Miltefosine Affects Lipid Metabolism in Leishmania donovani Promastigotes

Miltefosine Affects Lipid Metabolism in Leishmania donovani Promastigotes

Alteration of Fatty Acid and Sterol Metabolism in Miltefosine-Resistant Leishmania donovani Promastigotes and Consequences for Drug-Membrane Interactions

Thematic review series: Lipid Posttranslational Modifications. Fighting parasitic disease by blocking protein farnesylation

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