Sialic Acid Glycobiology Unveils Trypanosoma cruzi Trypomastigote Membrane Physiology

Trypanosoma cruzi, the flagellate protozoan agent of Chagas disease or American trypanosomiasis, is unable to synthesize sialic acids de novo. Mucins and trans-sialidase (TS) are substrate and enzyme, respectively, of the glycobiological system that scavenges sialic acid from the host in a crucial i...

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Publicado:	2016
Materias:	glycosylphosphatidylinositol anchored protein mucin phosphatidylinositol diacylglycerol lyase sialic acid sialidase glycoprotein n acetylneuraminic acid trans-sialidase affinity chromatography Article atomic force microscopy cell migration cell proliferation Chagas disease controlled study enzyme activity glycobiology immunofluorescence microscopy mass spectrometry membrane fluidity membrane microparticle nonhuman parasite survival parasite virulence protein expression protein purification sialylation transmission electron microscopy Trypanosoma cruzi trypomastigote ultracentrifugation Western blotting animal Bagg albino mouse disease model fluorescence microscopy host parasite interaction image processing metabolism microscopy mouse parasitology pathogenicity physiology procedures virulence Animals Cell-Derived Microparticles Chagas Disease Disease Models, Animal Glycoproteins Host-Parasite Interactions Image Processing, Computer-Assisted Mass Spectrometry Mice Mice, Inbred BALB C Microscopy Microscopy, Fluorescence Mucins N-Acetylneuraminic Acid Neuraminidase Virulence
Acceso en línea:	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_15537366_v12_n4_p_Lantos http://hdl.handle.net/20.500.12110/paper_15537366_v12_n4_p_Lantos
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

id	paper:paper_15537366_v12_n4_p_Lantos
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spelling	paper:paper_15537366_v12_n4_p_Lantos2023-06-08T16:23:10Z Sialic Acid Glycobiology Unveils Trypanosoma cruzi Trypomastigote Membrane Physiology glycosylphosphatidylinositol anchored protein mucin phosphatidylinositol diacylglycerol lyase sialic acid sialidase glycoprotein mucin n acetylneuraminic acid sialidase trans-sialidase affinity chromatography Article atomic force microscopy cell migration cell proliferation Chagas disease controlled study enzyme activity glycobiology immunofluorescence microscopy mass spectrometry membrane fluidity membrane microparticle nonhuman parasite survival parasite virulence protein expression protein purification sialylation transmission electron microscopy Trypanosoma cruzi trypomastigote ultracentrifugation Western blotting animal Bagg albino mouse Chagas disease disease model fluorescence microscopy host parasite interaction image processing metabolism microscopy mouse parasitology pathogenicity physiology procedures Trypanosoma cruzi virulence Animals Cell-Derived Microparticles Chagas Disease Disease Models, Animal Glycoproteins Host-Parasite Interactions Image Processing, Computer-Assisted Mass Spectrometry Mice Mice, Inbred BALB C Microscopy Microscopy, Fluorescence Mucins N-Acetylneuraminic Acid Neuraminidase Trypanosoma cruzi Virulence Trypanosoma cruzi, the flagellate protozoan agent of Chagas disease or American trypanosomiasis, is unable to synthesize sialic acids de novo. Mucins and trans-sialidase (TS) are substrate and enzyme, respectively, of the glycobiological system that scavenges sialic acid from the host in a crucial interplay for T. cruzi life cycle. The acquisition of the sialyl residue allows the parasite to avoid lysis by serum factors and to interact with the host cell. A major drawback to studying the sialylation kinetics and turnover of the trypomastigote glycoconjugates is the difficulty to identify and follow the recently acquired sialyl residues. To tackle this issue, we followed an unnatural sugar approach as bioorthogonal chemical reporters, where the use of azidosialyl residues allowed identifying the acquired sugar. Advanced microscopy techniques, together with biochemical methods, were used to study the trypomastigote membrane from its glycobiological perspective. Main sialyl acceptors were identified as mucins by biochemical procedures and protein markers. Together with determining their shedding and turnover rates, we also report that several membrane proteins, including TS and its substrates, both glycosylphosphatidylinositol-anchored proteins, are separately distributed on parasite surface and contained in different and highly stable membrane microdomains. Notably, labeling for α(1,3)Galactosyl residues only partially colocalize with sialylated mucins, indicating that two species of glycosylated mucins do exist, which are segregated at the parasite surface. Moreover, sialylated mucins were included in lipid-raft-domains, whereas TS molecules are not. The location of the surface-anchored TS resulted too far off as to be capable to sialylate mucins, a role played by the shed TS instead. Phosphatidylinositol-phospholipase-C activity is actually not present in trypomastigotes. Therefore, shedding of TS occurs via microvesicles instead of as a fully soluble form. © 2016 Lantos et al. 2016 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_15537366_v12_n4_p_Lantos http://hdl.handle.net/20.500.12110/paper_15537366_v12_n4_p_Lantos
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-134
collection	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic	glycosylphosphatidylinositol anchored protein mucin phosphatidylinositol diacylglycerol lyase sialic acid sialidase glycoprotein mucin n acetylneuraminic acid sialidase trans-sialidase affinity chromatography Article atomic force microscopy cell migration cell proliferation Chagas disease controlled study enzyme activity glycobiology immunofluorescence microscopy mass spectrometry membrane fluidity membrane microparticle nonhuman parasite survival parasite virulence protein expression protein purification sialylation transmission electron microscopy Trypanosoma cruzi trypomastigote ultracentrifugation Western blotting animal Bagg albino mouse Chagas disease disease model fluorescence microscopy host parasite interaction image processing metabolism microscopy mouse parasitology pathogenicity physiology procedures Trypanosoma cruzi virulence Animals Cell-Derived Microparticles Chagas Disease Disease Models, Animal Glycoproteins Host-Parasite Interactions Image Processing, Computer-Assisted Mass Spectrometry Mice Mice, Inbred BALB C Microscopy Microscopy, Fluorescence Mucins N-Acetylneuraminic Acid Neuraminidase Trypanosoma cruzi Virulence
spellingShingle	glycosylphosphatidylinositol anchored protein mucin phosphatidylinositol diacylglycerol lyase sialic acid sialidase glycoprotein mucin n acetylneuraminic acid sialidase trans-sialidase affinity chromatography Article atomic force microscopy cell migration cell proliferation Chagas disease controlled study enzyme activity glycobiology immunofluorescence microscopy mass spectrometry membrane fluidity membrane microparticle nonhuman parasite survival parasite virulence protein expression protein purification sialylation transmission electron microscopy Trypanosoma cruzi trypomastigote ultracentrifugation Western blotting animal Bagg albino mouse Chagas disease disease model fluorescence microscopy host parasite interaction image processing metabolism microscopy mouse parasitology pathogenicity physiology procedures Trypanosoma cruzi virulence Animals Cell-Derived Microparticles Chagas Disease Disease Models, Animal Glycoproteins Host-Parasite Interactions Image Processing, Computer-Assisted Mass Spectrometry Mice Mice, Inbred BALB C Microscopy Microscopy, Fluorescence Mucins N-Acetylneuraminic Acid Neuraminidase Trypanosoma cruzi Virulence Sialic Acid Glycobiology Unveils Trypanosoma cruzi Trypomastigote Membrane Physiology
topic_facet	glycosylphosphatidylinositol anchored protein mucin phosphatidylinositol diacylglycerol lyase sialic acid sialidase glycoprotein mucin n acetylneuraminic acid sialidase trans-sialidase affinity chromatography Article atomic force microscopy cell migration cell proliferation Chagas disease controlled study enzyme activity glycobiology immunofluorescence microscopy mass spectrometry membrane fluidity membrane microparticle nonhuman parasite survival parasite virulence protein expression protein purification sialylation transmission electron microscopy Trypanosoma cruzi trypomastigote ultracentrifugation Western blotting animal Bagg albino mouse Chagas disease disease model fluorescence microscopy host parasite interaction image processing metabolism microscopy mouse parasitology pathogenicity physiology procedures Trypanosoma cruzi virulence Animals Cell-Derived Microparticles Chagas Disease Disease Models, Animal Glycoproteins Host-Parasite Interactions Image Processing, Computer-Assisted Mass Spectrometry Mice Mice, Inbred BALB C Microscopy Microscopy, Fluorescence Mucins N-Acetylneuraminic Acid Neuraminidase Trypanosoma cruzi Virulence
description	Trypanosoma cruzi, the flagellate protozoan agent of Chagas disease or American trypanosomiasis, is unable to synthesize sialic acids de novo. Mucins and trans-sialidase (TS) are substrate and enzyme, respectively, of the glycobiological system that scavenges sialic acid from the host in a crucial interplay for T. cruzi life cycle. The acquisition of the sialyl residue allows the parasite to avoid lysis by serum factors and to interact with the host cell. A major drawback to studying the sialylation kinetics and turnover of the trypomastigote glycoconjugates is the difficulty to identify and follow the recently acquired sialyl residues. To tackle this issue, we followed an unnatural sugar approach as bioorthogonal chemical reporters, where the use of azidosialyl residues allowed identifying the acquired sugar. Advanced microscopy techniques, together with biochemical methods, were used to study the trypomastigote membrane from its glycobiological perspective. Main sialyl acceptors were identified as mucins by biochemical procedures and protein markers. Together with determining their shedding and turnover rates, we also report that several membrane proteins, including TS and its substrates, both glycosylphosphatidylinositol-anchored proteins, are separately distributed on parasite surface and contained in different and highly stable membrane microdomains. Notably, labeling for α(1,3)Galactosyl residues only partially colocalize with sialylated mucins, indicating that two species of glycosylated mucins do exist, which are segregated at the parasite surface. Moreover, sialylated mucins were included in lipid-raft-domains, whereas TS molecules are not. The location of the surface-anchored TS resulted too far off as to be capable to sialylate mucins, a role played by the shed TS instead. Phosphatidylinositol-phospholipase-C activity is actually not present in trypomastigotes. Therefore, shedding of TS occurs via microvesicles instead of as a fully soluble form. © 2016 Lantos et al.
title	Sialic Acid Glycobiology Unveils Trypanosoma cruzi Trypomastigote Membrane Physiology
title_short	Sialic Acid Glycobiology Unveils Trypanosoma cruzi Trypomastigote Membrane Physiology
title_full	Sialic Acid Glycobiology Unveils Trypanosoma cruzi Trypomastigote Membrane Physiology
title_fullStr	Sialic Acid Glycobiology Unveils Trypanosoma cruzi Trypomastigote Membrane Physiology
title_full_unstemmed	Sialic Acid Glycobiology Unveils Trypanosoma cruzi Trypomastigote Membrane Physiology
title_sort	sialic acid glycobiology unveils trypanosoma cruzi trypomastigote membrane physiology
publishDate	2016
url	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_15537366_v12_n4_p_Lantos http://hdl.handle.net/20.500.12110/paper_15537366_v12_n4_p_Lantos
_version_	1768545848592957440

Sialic Acid Glycobiology Unveils Trypanosoma cruzi Trypomastigote Membrane Physiology

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