Characterization of the cell adhesion site of Trypanosoma cruzi metacyclic stage surface glycoprotein gp82

Detalhes bibliográficos
Autor(a) principal: Manque, Patricio M. [UNIFESP]
Data de Publicação: 2000
Outros Autores: Eichinger, Daniel, Juliano, Maria Aparecida [UNIFESP], Juliano, Luiz [UNIFESP], Araya, Jorge E. [UNIFESP], Yoshida, Nobuko [UNIFESP]
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da UNIFESP
Texto Completo: http://dx.doi.org/10.1128/IAI.68.2.478-484.2000
http://repositorio.unifesp.br/handle/11600/26241
Resumo: The surface glycoprotein gp82, expressed in the insect-stage metacyclic trypomastigotes of Trypanosoma cruzi, has been implicated in mammalian cell invasion. Here we have characterized the cell adhesion site of gp82 by using recombinant proteins and synthetic peptides based on gp82. the recombinant protein Del-4/8, lacking 65 amino acids of gp82 central domain (at positions 257 to 321), was virtually devoid of cell-binding activity and lacked the ability to inhibit parasite invasion, in contrast to J18, the construct containing the full-length gp82 sequence (amino acids 1 to 516), Constructs with shorter deletions, i.e., Del-4 (deleted from 257 to 271) and Del-8 (deleted from 293 to 321), bound to target cells to a significantly lesser degree than did J18. the sites deleted in recombinant proteins Del-4 and Del-8 contained acidic amino acids critical for cell adhesion. Thus, the cell-binding capacity of protein Del-E/D, lacking the glutamic acid (259/260) and aspartic acid (303/304) pairs, was negligible, as was its capacity to inhibit parasite internalization. of a set of synthetic peptides spanning the gp82 central domain, a 22-mer hybrid peptide, p4/8, formed by two noncontiguous sequences (at positions 257 to 273 and 302 to 306) and containing the four acidic residues, competed with the binding of J18 protein to target cells and significantly inhibited (similar to 60%) the penetration of parasites. This peptide, generated by the juxtaposition of sequences that are separated by a hydrophobic stretch in the linear molecule, appears to be mimicking a conformation-dependent cell-binding site of gp82, Experiments of antibody competition with a set of 20-mer overlapping peptides mapped the epitope for 3F6, a monoclonal antibody directed to gp82 that inhibits parasite invasion, to the sequence represented by peptide p3 (244 to 263), which has a partial overlap with the cell adhesion site.
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spelling Characterization of the cell adhesion site of Trypanosoma cruzi metacyclic stage surface glycoprotein gp82The surface glycoprotein gp82, expressed in the insect-stage metacyclic trypomastigotes of Trypanosoma cruzi, has been implicated in mammalian cell invasion. Here we have characterized the cell adhesion site of gp82 by using recombinant proteins and synthetic peptides based on gp82. the recombinant protein Del-4/8, lacking 65 amino acids of gp82 central domain (at positions 257 to 321), was virtually devoid of cell-binding activity and lacked the ability to inhibit parasite invasion, in contrast to J18, the construct containing the full-length gp82 sequence (amino acids 1 to 516), Constructs with shorter deletions, i.e., Del-4 (deleted from 257 to 271) and Del-8 (deleted from 293 to 321), bound to target cells to a significantly lesser degree than did J18. the sites deleted in recombinant proteins Del-4 and Del-8 contained acidic amino acids critical for cell adhesion. Thus, the cell-binding capacity of protein Del-E/D, lacking the glutamic acid (259/260) and aspartic acid (303/304) pairs, was negligible, as was its capacity to inhibit parasite internalization. of a set of synthetic peptides spanning the gp82 central domain, a 22-mer hybrid peptide, p4/8, formed by two noncontiguous sequences (at positions 257 to 273 and 302 to 306) and containing the four acidic residues, competed with the binding of J18 protein to target cells and significantly inhibited (similar to 60%) the penetration of parasites. This peptide, generated by the juxtaposition of sequences that are separated by a hydrophobic stretch in the linear molecule, appears to be mimicking a conformation-dependent cell-binding site of gp82, Experiments of antibody competition with a set of 20-mer overlapping peptides mapped the epitope for 3F6, a monoclonal antibody directed to gp82 that inhibits parasite invasion, to the sequence represented by peptide p3 (244 to 263), which has a partial overlap with the cell adhesion site.Universidade Federal de São Paulo, Escola Paulista Med, Dept Microbiol Imunol & Parasitol, BR-04023062 São Paulo, BrazilUniversidade Federal de São Paulo, Escola Paulista Med, Dept Biofis, BR-04023062 São Paulo, BrazilNYU, Sch Med, Dept Med & Mol Parasitol, New York, NY USAUniversidade Federal de São Paulo, Escola Paulista Med, Dept Microbiol Imunol & Parasitol, BR-04023062 São Paulo, BrazilUniversidade Federal de São Paulo, Escola Paulista Med, Dept Biofis, BR-04023062 São Paulo, BrazilWeb of ScienceAmer Soc MicrobiologyUniversidade Federal de São Paulo (UNIFESP)NYUManque, Patricio M. [UNIFESP]Eichinger, DanielJuliano, Maria Aparecida [UNIFESP]Juliano, Luiz [UNIFESP]Araya, Jorge E. [UNIFESP]Yoshida, Nobuko [UNIFESP]2016-01-24T12:31:00Z2016-01-24T12:31:00Z2000-02-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersion478-484application/pdfhttp://dx.doi.org/10.1128/IAI.68.2.478-484.2000Infection and Immunity. Washington: Amer Soc Microbiology, v. 68, n. 2, p. 478-484, 2000.10.1128/IAI.68.2.478-484.2000WOS000084842000008.pdf0019-9567http://repositorio.unifesp.br/handle/11600/26241WOS:000084842000008engInfection and Immunityinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UNIFESPinstname:Universidade Federal de São Paulo (UNIFESP)instacron:UNIFESP2024-08-07T05:25:30Zoai:repositorio.unifesp.br/:11600/26241Repositório InstitucionalPUBhttp://www.repositorio.unifesp.br/oai/requestbiblioteca.csp@unifesp.bropendoar:34652024-08-07T05:25:30Repositório Institucional da UNIFESP - Universidade Federal de São Paulo (UNIFESP)false
dc.title.none.fl_str_mv Characterization of the cell adhesion site of Trypanosoma cruzi metacyclic stage surface glycoprotein gp82
title Characterization of the cell adhesion site of Trypanosoma cruzi metacyclic stage surface glycoprotein gp82
spellingShingle Characterization of the cell adhesion site of Trypanosoma cruzi metacyclic stage surface glycoprotein gp82
Manque, Patricio M. [UNIFESP]
title_short Characterization of the cell adhesion site of Trypanosoma cruzi metacyclic stage surface glycoprotein gp82
title_full Characterization of the cell adhesion site of Trypanosoma cruzi metacyclic stage surface glycoprotein gp82
title_fullStr Characterization of the cell adhesion site of Trypanosoma cruzi metacyclic stage surface glycoprotein gp82
title_full_unstemmed Characterization of the cell adhesion site of Trypanosoma cruzi metacyclic stage surface glycoprotein gp82
title_sort Characterization of the cell adhesion site of Trypanosoma cruzi metacyclic stage surface glycoprotein gp82
author Manque, Patricio M. [UNIFESP]
author_facet Manque, Patricio M. [UNIFESP]
Eichinger, Daniel
Juliano, Maria Aparecida [UNIFESP]
Juliano, Luiz [UNIFESP]
Araya, Jorge E. [UNIFESP]
Yoshida, Nobuko [UNIFESP]
author_role author
author2 Eichinger, Daniel
Juliano, Maria Aparecida [UNIFESP]
Juliano, Luiz [UNIFESP]
Araya, Jorge E. [UNIFESP]
Yoshida, Nobuko [UNIFESP]
author2_role author
author
author
author
author
dc.contributor.none.fl_str_mv Universidade Federal de São Paulo (UNIFESP)
NYU
dc.contributor.author.fl_str_mv Manque, Patricio M. [UNIFESP]
Eichinger, Daniel
Juliano, Maria Aparecida [UNIFESP]
Juliano, Luiz [UNIFESP]
Araya, Jorge E. [UNIFESP]
Yoshida, Nobuko [UNIFESP]
description The surface glycoprotein gp82, expressed in the insect-stage metacyclic trypomastigotes of Trypanosoma cruzi, has been implicated in mammalian cell invasion. Here we have characterized the cell adhesion site of gp82 by using recombinant proteins and synthetic peptides based on gp82. the recombinant protein Del-4/8, lacking 65 amino acids of gp82 central domain (at positions 257 to 321), was virtually devoid of cell-binding activity and lacked the ability to inhibit parasite invasion, in contrast to J18, the construct containing the full-length gp82 sequence (amino acids 1 to 516), Constructs with shorter deletions, i.e., Del-4 (deleted from 257 to 271) and Del-8 (deleted from 293 to 321), bound to target cells to a significantly lesser degree than did J18. the sites deleted in recombinant proteins Del-4 and Del-8 contained acidic amino acids critical for cell adhesion. Thus, the cell-binding capacity of protein Del-E/D, lacking the glutamic acid (259/260) and aspartic acid (303/304) pairs, was negligible, as was its capacity to inhibit parasite internalization. of a set of synthetic peptides spanning the gp82 central domain, a 22-mer hybrid peptide, p4/8, formed by two noncontiguous sequences (at positions 257 to 273 and 302 to 306) and containing the four acidic residues, competed with the binding of J18 protein to target cells and significantly inhibited (similar to 60%) the penetration of parasites. This peptide, generated by the juxtaposition of sequences that are separated by a hydrophobic stretch in the linear molecule, appears to be mimicking a conformation-dependent cell-binding site of gp82, Experiments of antibody competition with a set of 20-mer overlapping peptides mapped the epitope for 3F6, a monoclonal antibody directed to gp82 that inhibits parasite invasion, to the sequence represented by peptide p3 (244 to 263), which has a partial overlap with the cell adhesion site.
publishDate 2000
dc.date.none.fl_str_mv 2000-02-01
2016-01-24T12:31:00Z
2016-01-24T12:31:00Z
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://dx.doi.org/10.1128/IAI.68.2.478-484.2000
Infection and Immunity. Washington: Amer Soc Microbiology, v. 68, n. 2, p. 478-484, 2000.
10.1128/IAI.68.2.478-484.2000
WOS000084842000008.pdf
0019-9567
http://repositorio.unifesp.br/handle/11600/26241
WOS:000084842000008
url http://dx.doi.org/10.1128/IAI.68.2.478-484.2000
http://repositorio.unifesp.br/handle/11600/26241
identifier_str_mv Infection and Immunity. Washington: Amer Soc Microbiology, v. 68, n. 2, p. 478-484, 2000.
10.1128/IAI.68.2.478-484.2000
WOS000084842000008.pdf
0019-9567
WOS:000084842000008
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv Infection and Immunity
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv 478-484
application/pdf
dc.publisher.none.fl_str_mv Amer Soc Microbiology
publisher.none.fl_str_mv Amer Soc Microbiology
dc.source.none.fl_str_mv reponame:Repositório Institucional da UNIFESP
instname:Universidade Federal de São Paulo (UNIFESP)
instacron:UNIFESP
instname_str Universidade Federal de São Paulo (UNIFESP)
instacron_str UNIFESP
institution UNIFESP
reponame_str Repositório Institucional da UNIFESP
collection Repositório Institucional da UNIFESP
repository.name.fl_str_mv Repositório Institucional da UNIFESP - Universidade Federal de São Paulo (UNIFESP)
repository.mail.fl_str_mv biblioteca.csp@unifesp.br
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