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Isolation and characterization of a reservosome fraction from Trypanosoma cruzi

Cunha-e-Silva, Narcisa L ; Atella, Geórgia C ; Porto-Carreiro, Isabel A ; Morgado-Diaz, Jose A ; Pereira, Miria G ; de Souza, Wanderley

FEMS microbiology letters, 2002-08, Vol.214 (1), p.7-12 [Periódico revisado por pares]

Oxford, UK: Blackwell Publishing Ltd

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Título:
Isolation and characterization of a reservosome fraction from Trypanosoma cruzi
Autor: Cunha-e-Silva, Narcisa L ; Atella, Geórgia C ; Porto-Carreiro, Isabel A ; Morgado-Diaz, Jose A ; Pereira, Miria G ; de Souza, Wanderley
Assuntos: Animals ; Biochemistry. Physiology. Immunology. Molecular biology ; Biological and medical sciences ; Cell fractionation ; Centrifugation ; Centrifugation, Density Gradient - methods ; Cholesterol ; Cruzipain ; Cysteine proteinase ; Cytochrome ; Cytochrome c ; Cytochromes ; Endocytosis ; Endosomes - chemistry ; Endosomes - ultrastructure ; Epimastigotes ; Ergosterol ; Esterification ; Experimental protozoal diseases and models ; Fundamental and applied biological sciences. Psychology ; Gel electrophoresis ; Gels ; Infectious diseases ; Lecithin ; Lipids ; Lipids - analysis ; Markers ; Medical sciences ; Membranes ; Microbiology ; Microscopy, Electron ; Mitochondria ; Organelles ; Parasitic diseases ; Phosphatidylcholine ; Phosphatidylethanolamine ; Phospholipids ; Protease ; Protease inhibitors ; Proteinase ; Proteinase inhibitors ; Proteins ; Proteins - analysis ; Proteolysis ; Protozoa ; Protozoal diseases ; Pyrophosphatase ; Reductase ; Reductases ; Reservosome ; Sodium lauryl sulfate ; Subcellular Fractions - chemistry ; Subcellular Fractions - ultrastructure ; Substrates ; Sucrose ; Sugar ; Transmission electron microscopy ; Trypanosoma cruzi ; Trypanosoma cruzi - growth & development ; Trypanosoma cruzi - ultrastructure
É parte de: FEMS microbiology letters, 2002-08, Vol.214 (1), p.7-12
Notas: ObjectType-Article-2
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Descrição: Abstract Reservosomes are acidic compartments present at the posterior region of epimastigote forms of Trypanosoma cruzi that store proteins and lipids. During metacyclogenesis, they consume their contents and disappear. Reservosomes are rich in cruzipain, the main proteolytic enzyme of this parasite. By centrifugation in a sucrose gradient, we have obtained a highly purified subcellular fraction containing reservosomes from 5-day-old Y strain epimastigotes. Transmission electron microscopy showed that the fraction contained well-preserved organelles. The protein profile of the organelle analyzed by SDS–PAGE depicted a wide range of protein bands, predominating those corresponding to a triplet of 60–51 kDa and a doublet of 25–23 kDa. Protease activity in substrate-containing gels, in the presence or absence of protease inhibitors, showed that cysteine proteinase is enriched and very active in the purified fraction. Enzymatic assays demonstrated the absence of pyrophosphatase, an acidocalcisome marker, and succinate cytochrome c reductase, a mitochondrial marker, although these enzymes were active in other regions of the purification sucrose gradient. Thin layer chromatographic neutral lipid analysis of purified reservosomes demonstrated that the organelle stores large amounts of ergosterol and esterified cholesterol. Phospholipid analysis indicated phosphatidylcholine and phosphatidylethanolamine as the major constituents of reservosome membranes.
Editor: Oxford, UK: Blackwell Publishing Ltd
Idioma: Inglês

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Isolation and characterization of a reservosome fraction from Trypanosoma cruzi

Cunha-e-Silva, Narcisa L ; Atella, Geórgia C ; Porto-Carreiro, Isabel A ; Morgado-Diaz, Jose A ; Pereira, Miria G ; de Souza, Wanderley

Oxford, UK: Blackwell Publishing Ltd

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