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An evaluation of lipid metabolism in the insect trypanosomatid Herpetomonas muscarum uncovers a pathway for the uptake of extracellular insect lipoproteins
Authors:George Kluck  Karla C Régis  Nuccia NT De Cicco  Lívia Silva-Cardoso  Miria G Pereira  Patrícia Fampa  Alessandra C Chagas-Lima  Alexandre Romeiro  Narcisa L Cunha-Silva  Georgia C Atella
Institution:1. Instituto de Bioquímica Médica Leopoldo de Meis, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ RJ 21.941-902, Brazil;2. Instituto Nacional de Ciência e Tecnologia em Entomologia Molecular, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ RJ 21.941-902, Brazil;3. Departamento de Biologia Animal, Instituto de Biologia, Universidade Federal Rural do Rio de Janeiro, Seropédica, RJ RJ 23.890-000, Brazil;4. Laboratório de Ultraestrutura Celular Hertha Meyer, Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ RJ 21.941-902, Brazil
Abstract:Lipid uptake and metabolism by trypanosomatid parasites from vertebrate host blood have been well established in the literature. However, there is a lack of knowledge regarding the same aspects concerning the parasites that cross the hemolymph of their invertebrate hosts. We have investigated the lipid composition and metabolism of the insect trypanosomatid Herpetomonas muscarum by 3H- palmitic acid and phosphate (32Pi) and the parasite interaction with Lipophorin (Lp) the main lipid carrying protein of insect hemolymph. Gas chromatography-mass spectrometry (GC–MS) analyses were used to identify the fatty acids and sterols composition of H.muscarum. Furthermore, we investigated the Lp binding site in the plasma membrane of parasite by Immunolocalization. We showed that H. muscarum incorporated 3H-palmitic acid and inorganic phosphate (32Pi) which were readily used as precursor molecules of lipid biosynthetic pathways. Furthermore, H. muscarum was able to take up both protein and lipid moieties of Lp which could be used as nutrient sources. Moreover, we have also demonstrated for the first time the presence of a Lp binding site in the membrane of a parasite. Such results point out the role of describing the metabolic pathways of trypanosomatids in order to provide a better understanding of parasite-host interaction peculiarities. Such studies may enhance the potential form the identification of novel chemotherapeutic targets in harmful parasites.
Keywords:Lipid metabolism  Lipophorin  Insect trypanosomatid  Parasite-vector insect interaction  Sterols  Amu  Atomic mass units  CHO  Cholesterol  CHOE  Esterified cholesterol  DAG  Diacylglycerol  Lp  Lipophorin  125  Radiolabeled Lipophorin  32  Radioactive inorganic phosphate  3  Radioactive Fatty acid  FA  Fatty acid  FA-Bodipy  Palmitic acid Bodipy  IPC  Inositolphosphorylceramide  LPC  Lysophosphatidylcholine  MAG  Monoacylglycerol  ND  Not determined  NL  Neutral lipids  PA  Phosphatidic acid  PC  Phosphatidylcholine  PE  Phosphatidylethanolamine  PI  Phosphatidylinositol  PL  Phospholipids  PS  Phosphatidylserine  ST  Sterols  TAG  Triacylglycerol  TLC  Thin-layer chromatography
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