Structural analysis of the N-glycans of the major cysteine proteinase of Trypanosoma cruzi: Identification of sulfated high-mannose type oligosaccharides
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 a...
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todo:paper_1742464X_v272_n15_p3803_Barboza2023-10-03T16:30:19Z Structural analysis of the N-glycans of the major cysteine proteinase of Trypanosoma cruzi: Identification of sulfated high-mannose type oligosaccharides Barboza, M. Duschak, V.G. Fukuyama, Y. Nonami, H. Erra-Balsells, R. Cazzulo, J.J. Couto, A.S. 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 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. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_1742464X_v272_n15_p3803_Barboza |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
| collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
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 |
| spellingShingle |
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 Barboza, M. Duschak, V.G. Fukuyama, Y. Nonami, H. Erra-Balsells, R. Cazzulo, J.J. Couto, A.S. Structural analysis of the N-glycans of the major cysteine proteinase of Trypanosoma cruzi: Identification of sulfated high-mannose type oligosaccharides |
| topic_facet |
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 |
| description |
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. |
| format |
JOUR |
| author |
Barboza, M. Duschak, V.G. Fukuyama, Y. Nonami, H. Erra-Balsells, R. Cazzulo, J.J. Couto, A.S. |
| author_facet |
Barboza, M. Duschak, V.G. Fukuyama, Y. Nonami, H. Erra-Balsells, R. Cazzulo, J.J. Couto, A.S. |
| author_sort |
Barboza, M. |
| title |
Structural analysis of the N-glycans of the major cysteine proteinase of Trypanosoma cruzi: Identification of sulfated high-mannose type oligosaccharides |
| title_short |
Structural analysis of the N-glycans of the major cysteine proteinase of Trypanosoma cruzi: Identification of sulfated high-mannose type oligosaccharides |
| title_full |
Structural analysis of the N-glycans of the major cysteine proteinase of Trypanosoma cruzi: Identification of sulfated high-mannose type oligosaccharides |
| title_fullStr |
Structural analysis of the N-glycans of the major cysteine proteinase of Trypanosoma cruzi: Identification of sulfated high-mannose type oligosaccharides |
| title_full_unstemmed |
Structural analysis of the N-glycans of the major cysteine proteinase of Trypanosoma cruzi: Identification of sulfated high-mannose type oligosaccharides |
| title_sort |
structural analysis of the n-glycans of the major cysteine proteinase of trypanosoma cruzi: identification of sulfated high-mannose type oligosaccharides |
| url |
http://hdl.handle.net/20.500.12110/paper_1742464X_v272_n15_p3803_Barboza |
| work_keys_str_mv |
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