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Abstract:

Trypanosoma cruzi, the parasitic protozoan that causes Chagas disease, contains a major cysteine proteinase, cruzipain. This lysosomal enzyme bears an unusual C-terminal extension that contains a number of post-translational modifications, and most antibodies in natural and experimental infections are directed against it. In this report we took advantage of UV-MALDI-TOF mass spectrometry in conjunction with peptide N-glycosidase F deglycosylation and high performance anion exchange chromatography analysis to address the structure of the N-linked oligosaccharides present in this domain. The UV-MALDI-TOF MS analysis in the negative-ion mode, using nor-harmane as matrix, allowed us to determine a new striking feature in cruzipain: sulfated high-mannose type oligosaccharides. Sulfated GlcNAc2Man3 to GlcNAc 2Man9 species were identified. In accordance, after chemical or enzymatic desulfation, the corresponding signals disappeared. In addition, by UV-MALDI-TOF MS analysis (a) a main population of high-mannose type oligosaccharides was shown in the positive-ion mode, (b) lactosaminic glycans were also identified, among them, structures corresponding to monosialylated species were detected, and (c) as an interesting fact a fucosylated oligosaccharide was also detected. The presence of the deoxy sugar was further confirmed by high performance anion exchange chromatography. In conclusion, the total number of oligosaccharides occurring in cruzipain was shown to be much higher than previous estimates. This constitutes the first report on the presence of sulfated glycoproteins in Trypanosomatids. © 2005 FEBS.

Registro:

Documento: Artículo
Título:Structural analysis of the N-glycans of the major cysteine proteinase of Trypanosoma cruzi: Identification of sulfated high-mannose type oligosaccharides
Autor:Barboza, M.; Duschak, V.G.; Fukuyama, Y.; Nonami, H.; Erra-Balsells, R.; Cazzulo, J.J.; Couto, A.S.
Filiación:Instituto de Investigaciones Biotecnológicas, INTECH, Universidad Nacional de Gral, San Martin, Buenos Aires, Argentina
Instituto Nacional de Parasitología Dr. Mario Fatala Chabén, ANLIS, Ministerio de Salud y Ambiente, Buenos Aires, Argentina
College of Agriculture, Ehime University, Matsuyama, Japan
CIHIDECAR (CONICET), Departamento de Química Orgánica, Universidad de Buenos Aires, Buenos Aires, Argentina
CIHIDECAR (CONICET), Departamento de Química Orgánica, Universidad de Buenos Aires, CP 1428, Buenos Aires, Argentina
Koichi Tanaka Mass Spectrometry Research Laboratory, Shimadzu Corporation, 1 Nishinokyo-Kuwabaracho, Nakagyo-ku, Kyoto 604-8511, Japan
Palabras clave:Cruzipain; Nor-harmane; Sulfated oligosaccharides; Trypanosoma cruzi; UV-MALDI-TOF MS; cruzipain; cysteine proteinase; deoxysugar; glycan; glycosidase; mannose; oligosaccharide; anion exchange chromatography; article; carboxy terminal sequence; deglycosylation; fucosylation; matrix assisted laser desorption ionization time of flight mass spectrometry; nonhuman; priority journal; protein analysis; sialylation; structure analysis; sulfation; Trypanosoma cruzi; Animals; Cysteine Endopeptidases; Electrophoresis, Polyacrylamide Gel; Fucose; Mannose; Oligosaccharides, Branched-Chain; Silver Staining; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Time Factors; Trypanosoma cruzi; Protozoa; Trypanosoma cruzi; Trypanosomatidae
Año:2005
Volumen:272
Número:15
Página de inicio:3803
Página de fin:3815
DOI: http://dx.doi.org/10.1111/j.1742-4658.2005.04787.x
Título revista:FEBS Journal
Título revista abreviado:FEBS J.
ISSN:1742464X
CAS:cysteine proteinase, 37353-41-6; glycosidase, 9032-92-2; mannose, 31103-86-3, 3458-28-4; cruzipain, EC 3.4.22.-; Cysteine Endopeptidases, EC 3.4.22.-; Fucose, 3713-31-3; Mannose, 31103-86-3; Oligosaccharides, Branched-Chain
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_1742464X_v272_n15_p3803_Barboza

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Citas:

---------- APA ----------
Barboza, M., Duschak, V.G., Fukuyama, Y., Nonami, H., Erra-Balsells, R., Cazzulo, J.J. & Couto, A.S. (2005) . Structural analysis of the N-glycans of the major cysteine proteinase of Trypanosoma cruzi: Identification of sulfated high-mannose type oligosaccharides. FEBS Journal, 272(15), 3803-3815.
http://dx.doi.org/10.1111/j.1742-4658.2005.04787.x
---------- CHICAGO ----------
Barboza, M., Duschak, V.G., Fukuyama, Y., Nonami, H., Erra-Balsells, R., Cazzulo, J.J., et al. "Structural analysis of the N-glycans of the major cysteine proteinase of Trypanosoma cruzi: Identification of sulfated high-mannose type oligosaccharides" . FEBS Journal 272, no. 15 (2005) : 3803-3815.
http://dx.doi.org/10.1111/j.1742-4658.2005.04787.x
---------- MLA ----------
Barboza, M., Duschak, V.G., Fukuyama, Y., Nonami, H., Erra-Balsells, R., Cazzulo, J.J., et al. "Structural analysis of the N-glycans of the major cysteine proteinase of Trypanosoma cruzi: Identification of sulfated high-mannose type oligosaccharides" . FEBS Journal, vol. 272, no. 15, 2005, pp. 3803-3815.
http://dx.doi.org/10.1111/j.1742-4658.2005.04787.x
---------- VANCOUVER ----------
Barboza, M., Duschak, V.G., Fukuyama, Y., Nonami, H., Erra-Balsells, R., Cazzulo, J.J., et al. Structural analysis of the N-glycans of the major cysteine proteinase of Trypanosoma cruzi: Identification of sulfated high-mannose type oligosaccharides. FEBS J. 2005;272(15):3803-3815.
http://dx.doi.org/10.1111/j.1742-4658.2005.04787.x