An anionic synthetic sugar containing 6-SO3-NAcGlc mimics the sulfated cruzipain epitope that plays a central role in immune recognition
Cruzipain (Cz), the major cysteine proteinase of Trypanosoma cruzi, is a glycoprotein that contains sulfated high-mannose-type oligosaccharides. We have previously determined that these sulfate groups are targets of specific immune responses. In order to evaluate the structural requirements for anti...
Guardado en:
| Autores principales: | , , , , , , , , , , , , |
|---|---|
| Formato: | JOUR |
| Materias: | |
| Acceso en línea: | http://hdl.handle.net/20.500.12110/paper_1742464X_v279_n19_p3665_Couto |
| Aporte de: |
| id |
todo:paper_1742464X_v279_n19_p3665_Couto |
|---|---|
| record_format |
dspace |
| spelling |
todo:paper_1742464X_v279_n19_p3665_Couto2023-10-03T16:30:20Z An anionic synthetic sugar containing 6-SO3-NAcGlc mimics the sulfated cruzipain epitope that plays a central role in immune recognition Couto, A.S. Soprano, L.L. Landoni, M. Pourcelot, M. Acosta, D.M. Bultel, L. Parente, J. Ferrero, M.R. Barbier, M. Dussouy, C. Esteva, M.I. Kovensky, J. Duschak, V.G. cruzipain epitope glycoprotein sulfated GlcNAc Trypanosoma cruzi acetic acid derivative aprotinin bovine serum albumin cruzipain disaccharide immunoglobulin G inorganic salt monosaccharide n acetyl dextro glucosamine 6 sulfate sodium salt oligosaccharide sulfate unclassified drug adult animal experiment antibody production antigen binding antigen recognition article binding affinity carboxy terminal sequence carboxylation Chagas disease competitive inhibition controlled study enzyme modification enzyme structure esterification female human immunization immunoassay mouse nonhuman nucleotide sequence phosphorylation priority journal rabbit structure activity relation sulfation surface charge Trypanosoma cruzi Acetylglucosamine Adolescent Adult Animals Anions Case-Control Studies Chagas Disease Cysteine Endopeptidases Enzyme-Linked Immunosorbent Assay Epitopes Female Humans Immunoglobulin G Magnetic Resonance Spectroscopy Male Mice Mice, Inbred BALB C Middle Aged Oligosaccharides Rabbits Serologic Tests Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization Sulfates Trypanosoma cruzi Young Adult Oryctolagus cuniculus Trypanosoma cruzi Cruzipain (Cz), the major cysteine proteinase of Trypanosoma cruzi, is a glycoprotein that contains sulfated high-mannose-type oligosaccharides. We have previously determined that these sulfate groups are targets of specific immune responses. In order to evaluate the structural requirements for antibody recognition of Cz, a systematic structure-activity study of the chemical characteristics needed for antibody binding to the Cz sulfated epitope was performed by immunoassays. With this aim, different synthesized molecules were coupled to the proteins BSA and aprotinin and confronted with (a) mouse sera specific for Cz and its carboxy-terminal (C-T) domain, (b) antibodies raised in rabbits immunized with Cz and its C-terminal domain and (c) IgGs purified from human Chagas disease sera. Our results indicate that a glucosamine containing an esterifying sulfate group in position O-6 and an N-acetyl group was the preferred epitope for the immune recognition of sera specific for Cz and its C-T domain. Although to a minor extent, other anionic compounds bearing sulfate groups in different positions and number as well as different anionic charged groups including carboxylated or phosphorylated monosaccharides, disaccharides and oligosaccharides were recognized. In conclusion, we found that synthetic anionic sugar conjugates containing N-acetyl d-glucosamine-6-sulfate sodium salt (GlcNAc6S) competitively inhibit the binding of affinity purified rabbit anti-C-T IgG to the C-T extension of Cz. Extending these findings to the context of natural infection, immune assays performed with Chagas disease serum confirmed that the structure of synthetic GlcNAc6S mimics the N-glycan-linked sulfated epitope displayed in the C-T domain of Cz. A systematic study of the chemical characteristics needed for antibody binding to the cruzipain sulfated epitope was performed by immunoassays. Different molecules were synthesized, coupled to BSA/aprotinin, and confronted with rabbit/mice sera specific for Cz/C-T and purified IgGs from immune and human Chagas disease sera, demonstrating that synthetic GlcNAc6S mimics the N-glycan-linked-sulfated epitope displayed in the C-T domain of natural cruzipain. © 2012 The Authors Journal compilation © 2012 FEBS. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_1742464X_v279_n19_p3665_Couto |
| 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 epitope glycoprotein sulfated GlcNAc Trypanosoma cruzi acetic acid derivative aprotinin bovine serum albumin cruzipain disaccharide immunoglobulin G inorganic salt monosaccharide n acetyl dextro glucosamine 6 sulfate sodium salt oligosaccharide sulfate unclassified drug adult animal experiment antibody production antigen binding antigen recognition article binding affinity carboxy terminal sequence carboxylation Chagas disease competitive inhibition controlled study enzyme modification enzyme structure esterification female human immunization immunoassay mouse nonhuman nucleotide sequence phosphorylation priority journal rabbit structure activity relation sulfation surface charge Trypanosoma cruzi Acetylglucosamine Adolescent Adult Animals Anions Case-Control Studies Chagas Disease Cysteine Endopeptidases Enzyme-Linked Immunosorbent Assay Epitopes Female Humans Immunoglobulin G Magnetic Resonance Spectroscopy Male Mice Mice, Inbred BALB C Middle Aged Oligosaccharides Rabbits Serologic Tests Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization Sulfates Trypanosoma cruzi Young Adult Oryctolagus cuniculus Trypanosoma cruzi |
| spellingShingle |
cruzipain epitope glycoprotein sulfated GlcNAc Trypanosoma cruzi acetic acid derivative aprotinin bovine serum albumin cruzipain disaccharide immunoglobulin G inorganic salt monosaccharide n acetyl dextro glucosamine 6 sulfate sodium salt oligosaccharide sulfate unclassified drug adult animal experiment antibody production antigen binding antigen recognition article binding affinity carboxy terminal sequence carboxylation Chagas disease competitive inhibition controlled study enzyme modification enzyme structure esterification female human immunization immunoassay mouse nonhuman nucleotide sequence phosphorylation priority journal rabbit structure activity relation sulfation surface charge Trypanosoma cruzi Acetylglucosamine Adolescent Adult Animals Anions Case-Control Studies Chagas Disease Cysteine Endopeptidases Enzyme-Linked Immunosorbent Assay Epitopes Female Humans Immunoglobulin G Magnetic Resonance Spectroscopy Male Mice Mice, Inbred BALB C Middle Aged Oligosaccharides Rabbits Serologic Tests Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization Sulfates Trypanosoma cruzi Young Adult Oryctolagus cuniculus Trypanosoma cruzi Couto, A.S. Soprano, L.L. Landoni, M. Pourcelot, M. Acosta, D.M. Bultel, L. Parente, J. Ferrero, M.R. Barbier, M. Dussouy, C. Esteva, M.I. Kovensky, J. Duschak, V.G. An anionic synthetic sugar containing 6-SO3-NAcGlc mimics the sulfated cruzipain epitope that plays a central role in immune recognition |
| topic_facet |
cruzipain epitope glycoprotein sulfated GlcNAc Trypanosoma cruzi acetic acid derivative aprotinin bovine serum albumin cruzipain disaccharide immunoglobulin G inorganic salt monosaccharide n acetyl dextro glucosamine 6 sulfate sodium salt oligosaccharide sulfate unclassified drug adult animal experiment antibody production antigen binding antigen recognition article binding affinity carboxy terminal sequence carboxylation Chagas disease competitive inhibition controlled study enzyme modification enzyme structure esterification female human immunization immunoassay mouse nonhuman nucleotide sequence phosphorylation priority journal rabbit structure activity relation sulfation surface charge Trypanosoma cruzi Acetylglucosamine Adolescent Adult Animals Anions Case-Control Studies Chagas Disease Cysteine Endopeptidases Enzyme-Linked Immunosorbent Assay Epitopes Female Humans Immunoglobulin G Magnetic Resonance Spectroscopy Male Mice Mice, Inbred BALB C Middle Aged Oligosaccharides Rabbits Serologic Tests Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization Sulfates Trypanosoma cruzi Young Adult Oryctolagus cuniculus Trypanosoma cruzi |
| description |
Cruzipain (Cz), the major cysteine proteinase of Trypanosoma cruzi, is a glycoprotein that contains sulfated high-mannose-type oligosaccharides. We have previously determined that these sulfate groups are targets of specific immune responses. In order to evaluate the structural requirements for antibody recognition of Cz, a systematic structure-activity study of the chemical characteristics needed for antibody binding to the Cz sulfated epitope was performed by immunoassays. With this aim, different synthesized molecules were coupled to the proteins BSA and aprotinin and confronted with (a) mouse sera specific for Cz and its carboxy-terminal (C-T) domain, (b) antibodies raised in rabbits immunized with Cz and its C-terminal domain and (c) IgGs purified from human Chagas disease sera. Our results indicate that a glucosamine containing an esterifying sulfate group in position O-6 and an N-acetyl group was the preferred epitope for the immune recognition of sera specific for Cz and its C-T domain. Although to a minor extent, other anionic compounds bearing sulfate groups in different positions and number as well as different anionic charged groups including carboxylated or phosphorylated monosaccharides, disaccharides and oligosaccharides were recognized. In conclusion, we found that synthetic anionic sugar conjugates containing N-acetyl d-glucosamine-6-sulfate sodium salt (GlcNAc6S) competitively inhibit the binding of affinity purified rabbit anti-C-T IgG to the C-T extension of Cz. Extending these findings to the context of natural infection, immune assays performed with Chagas disease serum confirmed that the structure of synthetic GlcNAc6S mimics the N-glycan-linked sulfated epitope displayed in the C-T domain of Cz. A systematic study of the chemical characteristics needed for antibody binding to the cruzipain sulfated epitope was performed by immunoassays. Different molecules were synthesized, coupled to BSA/aprotinin, and confronted with rabbit/mice sera specific for Cz/C-T and purified IgGs from immune and human Chagas disease sera, demonstrating that synthetic GlcNAc6S mimics the N-glycan-linked-sulfated epitope displayed in the C-T domain of natural cruzipain. © 2012 The Authors Journal compilation © 2012 FEBS. |
| format |
JOUR |
| author |
Couto, A.S. Soprano, L.L. Landoni, M. Pourcelot, M. Acosta, D.M. Bultel, L. Parente, J. Ferrero, M.R. Barbier, M. Dussouy, C. Esteva, M.I. Kovensky, J. Duschak, V.G. |
| author_facet |
Couto, A.S. Soprano, L.L. Landoni, M. Pourcelot, M. Acosta, D.M. Bultel, L. Parente, J. Ferrero, M.R. Barbier, M. Dussouy, C. Esteva, M.I. Kovensky, J. Duschak, V.G. |
| author_sort |
Couto, A.S. |
| title |
An anionic synthetic sugar containing 6-SO3-NAcGlc mimics the sulfated cruzipain epitope that plays a central role in immune recognition |
| title_short |
An anionic synthetic sugar containing 6-SO3-NAcGlc mimics the sulfated cruzipain epitope that plays a central role in immune recognition |
| title_full |
An anionic synthetic sugar containing 6-SO3-NAcGlc mimics the sulfated cruzipain epitope that plays a central role in immune recognition |
| title_fullStr |
An anionic synthetic sugar containing 6-SO3-NAcGlc mimics the sulfated cruzipain epitope that plays a central role in immune recognition |
| title_full_unstemmed |
An anionic synthetic sugar containing 6-SO3-NAcGlc mimics the sulfated cruzipain epitope that plays a central role in immune recognition |
| title_sort |
anionic synthetic sugar containing 6-so3-nacglc mimics the sulfated cruzipain epitope that plays a central role in immune recognition |
| url |
http://hdl.handle.net/20.500.12110/paper_1742464X_v279_n19_p3665_Couto |
| work_keys_str_mv |
AT coutoas ananionicsyntheticsugarcontaining6so3nacglcmimicsthesulfatedcruzipainepitopethatplaysacentralroleinimmunerecognition AT sopranoll ananionicsyntheticsugarcontaining6so3nacglcmimicsthesulfatedcruzipainepitopethatplaysacentralroleinimmunerecognition AT landonim ananionicsyntheticsugarcontaining6so3nacglcmimicsthesulfatedcruzipainepitopethatplaysacentralroleinimmunerecognition AT pourcelotm ananionicsyntheticsugarcontaining6so3nacglcmimicsthesulfatedcruzipainepitopethatplaysacentralroleinimmunerecognition AT acostadm ananionicsyntheticsugarcontaining6so3nacglcmimicsthesulfatedcruzipainepitopethatplaysacentralroleinimmunerecognition AT bultell ananionicsyntheticsugarcontaining6so3nacglcmimicsthesulfatedcruzipainepitopethatplaysacentralroleinimmunerecognition AT parentej ananionicsyntheticsugarcontaining6so3nacglcmimicsthesulfatedcruzipainepitopethatplaysacentralroleinimmunerecognition AT ferreromr ananionicsyntheticsugarcontaining6so3nacglcmimicsthesulfatedcruzipainepitopethatplaysacentralroleinimmunerecognition AT barbierm ananionicsyntheticsugarcontaining6so3nacglcmimicsthesulfatedcruzipainepitopethatplaysacentralroleinimmunerecognition AT dussouyc ananionicsyntheticsugarcontaining6so3nacglcmimicsthesulfatedcruzipainepitopethatplaysacentralroleinimmunerecognition AT estevami ananionicsyntheticsugarcontaining6so3nacglcmimicsthesulfatedcruzipainepitopethatplaysacentralroleinimmunerecognition AT kovenskyj ananionicsyntheticsugarcontaining6so3nacglcmimicsthesulfatedcruzipainepitopethatplaysacentralroleinimmunerecognition AT duschakvg ananionicsyntheticsugarcontaining6so3nacglcmimicsthesulfatedcruzipainepitopethatplaysacentralroleinimmunerecognition AT coutoas anionicsyntheticsugarcontaining6so3nacglcmimicsthesulfatedcruzipainepitopethatplaysacentralroleinimmunerecognition AT sopranoll anionicsyntheticsugarcontaining6so3nacglcmimicsthesulfatedcruzipainepitopethatplaysacentralroleinimmunerecognition AT landonim anionicsyntheticsugarcontaining6so3nacglcmimicsthesulfatedcruzipainepitopethatplaysacentralroleinimmunerecognition AT pourcelotm anionicsyntheticsugarcontaining6so3nacglcmimicsthesulfatedcruzipainepitopethatplaysacentralroleinimmunerecognition AT acostadm anionicsyntheticsugarcontaining6so3nacglcmimicsthesulfatedcruzipainepitopethatplaysacentralroleinimmunerecognition AT bultell anionicsyntheticsugarcontaining6so3nacglcmimicsthesulfatedcruzipainepitopethatplaysacentralroleinimmunerecognition AT parentej anionicsyntheticsugarcontaining6so3nacglcmimicsthesulfatedcruzipainepitopethatplaysacentralroleinimmunerecognition AT ferreromr anionicsyntheticsugarcontaining6so3nacglcmimicsthesulfatedcruzipainepitopethatplaysacentralroleinimmunerecognition AT barbierm anionicsyntheticsugarcontaining6so3nacglcmimicsthesulfatedcruzipainepitopethatplaysacentralroleinimmunerecognition AT dussouyc anionicsyntheticsugarcontaining6so3nacglcmimicsthesulfatedcruzipainepitopethatplaysacentralroleinimmunerecognition AT estevami anionicsyntheticsugarcontaining6so3nacglcmimicsthesulfatedcruzipainepitopethatplaysacentralroleinimmunerecognition AT kovenskyj anionicsyntheticsugarcontaining6so3nacglcmimicsthesulfatedcruzipainepitopethatplaysacentralroleinimmunerecognition AT duschakvg anionicsyntheticsugarcontaining6so3nacglcmimicsthesulfatedcruzipainepitopethatplaysacentralroleinimmunerecognition |
| _version_ |
1782024897719959552 |