Identification of complete precursors for the glycosylphosphatidylinositol protein anchors of Trypanosoma cruzi
Autor(a) principal: | |
---|---|
Data de Publicação: | 1996 |
Outros Autores: | , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNIFESP |
Texto Completo: | http://repositorio.unifesp.br/11600/44205 http://doi.org/10.1074/jbc.271.28.16877 |
Resumo: | The survival of Trypanosoma cruzi, the causative agent of Chagas' disease, depends vitally on proteins and glycoconjugates that mediate the parasite/host interaction. Since most of these molecules are attached to the membrane by glycosylphosphatidylinositol (GPI), alternative means of chemotherapeutic intervention might emerge from GPI biosynthesis studies. The structure of the major 1G7 antigen GPI has been fully characterized by us (Guther, M. L. S., Cardoso de Almeida, M. L., Yoshida, N., and Ferguson, M. A. J. (1992) J. Biol. Chem. 267, 6820-6828; Heise, N., Cardoso de Almeida, M. L., and Ferguson, M. A. J. (1995) Mel. Biochem. Parasitol. 70, 71-84), and based on its properties we now report the complete precursor glycolipids predicted to be transferred to the nascent protein. Migrating closely to Trypanosoma brucei glycolipid A on TLC, such species, named glycolipids A-like 1 and A-like 2, were labeled with tritiated palmitic acid, myo-inositol, glucosamine, and mannose, but surprisingly only the less polar glycolipid A-like 1 incorporated ethanolamine. The predicted products following nitrous acid deamination and digestion with phospholipases A(2), C, and D confirmed their GPI nature. Evidence that they may represent the anchor transferred to the 1G7 antigen came from the following analyses: (i) alpha-mannosidase treatments indicated that only one mannose was amenable to removal; (ii) their lipid moiety was identified as sn-l-alkyl-a-acylglycerol due to their sensitivity to phospholipase A(2) (PLA(2)), mild base and by direct high performance TLC analysis of the corresponding benzoylated diradylglycerol components; and (iii) both glycolipids incorporated H-3-fatty acid only in the sn-2- and not in the sn-l-alkyl position as previously found in the GPI of the mature 1G7 antigen. Based on the differential [H-3]ethanolamine incorporation pattern and the recent report that an aminoethylphosphonic acid (AEP) replaces ethanolamine phosphate (EtNH(2)-PO4) in the GPI in epimastigote sialoglycoproteins (Previato, J. O., Jones, C., Xavier, M. T., Wait, R., Travassos, L. R., Parodi, A. J., and Mendonca-Previato, L. (1995) J. Biol. Chem. 270, 7241-7250) it is proposed that glycolipid A-like 2 contains AEP and A-like 1 EtNH(2)-PO4. In the in vitro cell-free system both glycolipids were synthesized simultaneously and do not seem to bear a precursor/product relationship. Among the various components synthesized in vitro a glycolipid C-like corresponding to a form of glycolipid A-like 1 acylated on the inositol was also characterized. Phenylmethylsulfonyl fluoride, an inhibitor known to block the addition of ethanolamine phosphate in T. brucei but not in mammalian cells, also inhibits the synthesis of glycolipids A like and C-like in T. cruzi, indicating that the putative trypanosome EtNH(2)-PO4/AEP transferase(s) might represent a potential target for chemotherapy. |
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Heise, Norton [UNIFESP]Raper, J.Buxbaum, L. U.Peranovich, TMSAlmeida, MLCUniversidade Federal de São Paulo (UNIFESP)JOHNS HOPKINS UNIV2018-06-15T17:53:05Z2018-06-15T17:53:05Z1996-07-12Journal Of Biological Chemistry. Bethesda: Amer Soc Biochemistry Molecular Biology Inc, v. 271, n. 28, p. 16877-16887, 1996.0021-9258http://repositorio.unifesp.br/11600/44205http://doi.org/10.1074/jbc.271.28.1687710.1074/jbc.271.28.16877WOS:A1996UX12600070The survival of Trypanosoma cruzi, the causative agent of Chagas' disease, depends vitally on proteins and glycoconjugates that mediate the parasite/host interaction. Since most of these molecules are attached to the membrane by glycosylphosphatidylinositol (GPI), alternative means of chemotherapeutic intervention might emerge from GPI biosynthesis studies. The structure of the major 1G7 antigen GPI has been fully characterized by us (Guther, M. L. S., Cardoso de Almeida, M. L., Yoshida, N., and Ferguson, M. A. J. (1992) J. Biol. Chem. 267, 6820-6828; Heise, N., Cardoso de Almeida, M. L., and Ferguson, M. A. J. (1995) Mel. Biochem. Parasitol. 70, 71-84), and based on its properties we now report the complete precursor glycolipids predicted to be transferred to the nascent protein. Migrating closely to Trypanosoma brucei glycolipid A on TLC, such species, named glycolipids A-like 1 and A-like 2, were labeled with tritiated palmitic acid, myo-inositol, glucosamine, and mannose, but surprisingly only the less polar glycolipid A-like 1 incorporated ethanolamine. The predicted products following nitrous acid deamination and digestion with phospholipases A(2), C, and D confirmed their GPI nature. Evidence that they may represent the anchor transferred to the 1G7 antigen came from the following analyses: (i) alpha-mannosidase treatments indicated that only one mannose was amenable to removal; (ii) their lipid moiety was identified as sn-l-alkyl-a-acylglycerol due to their sensitivity to phospholipase A(2) (PLA(2)), mild base and by direct high performance TLC analysis of the corresponding benzoylated diradylglycerol components; and (iii) both glycolipids incorporated H-3-fatty acid only in the sn-2- and not in the sn-l-alkyl position as previously found in the GPI of the mature 1G7 antigen. Based on the differential [H-3]ethanolamine incorporation pattern and the recent report that an aminoethylphosphonic acid (AEP) replaces ethanolamine phosphate (EtNH(2)-PO4) in the GPI in epimastigote sialoglycoproteins (Previato, J. O., Jones, C., Xavier, M. T., Wait, R., Travassos, L. R., Parodi, A. J., and Mendonca-Previato, L. (1995) J. Biol. Chem. 270, 7241-7250) it is proposed that glycolipid A-like 2 contains AEP and A-like 1 EtNH(2)-PO4. In the in vitro cell-free system both glycolipids were synthesized simultaneously and do not seem to bear a precursor/product relationship. Among the various components synthesized in vitro a glycolipid C-like corresponding to a form of glycolipid A-like 1 acylated on the inositol was also characterized. Phenylmethylsulfonyl fluoride, an inhibitor known to block the addition of ethanolamine phosphate in T. brucei but not in mammalian cells, also inhibits the synthesis of glycolipids A like and C-like in T. cruzi, indicating that the putative trypanosome EtNH(2)-PO4/AEP transferase(s) might represent a potential target for chemotherapy.UNIV FED SAO PAULO,ESCOLA PAULISTA MED,DEPT MICROBIOL IMMUNOL & PARASITOL,BR-04023062 SAO PAULO,SP,BRAZILJOHNS HOPKINS UNIV,SCH MED,DEPT BIOL CHEM,BALTIMORE,MD 21205UNIV FED SAO PAULO,ESCOLA PAULISTA MED,DEPT MICROBIOL IMMUNOL & PARASITOL,BR-04023062 SAO PAULO,SP,BRAZILWeb of Science16877-16887engAmer Soc Biochemistry Molecular Biology IncJournal Of Biological ChemistryIdentification of complete precursors for the glycosylphosphatidylinositol protein anchors of Trypanosoma cruziinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UNIFESPinstname:Universidade Federal de São Paulo (UNIFESP)instacron:UNIFESP11600/442052022-02-08 11:52:10.523metadata only accessoai:repositorio.unifesp.br:11600/44205Repositório InstitucionalPUBhttp://www.repositorio.unifesp.br/oai/requestopendoar:34652022-02-08T14:52:10Repositório Institucional da UNIFESP - Universidade Federal de São Paulo (UNIFESP)false |
dc.title.en.fl_str_mv |
Identification of complete precursors for the glycosylphosphatidylinositol protein anchors of Trypanosoma cruzi |
title |
Identification of complete precursors for the glycosylphosphatidylinositol protein anchors of Trypanosoma cruzi |
spellingShingle |
Identification of complete precursors for the glycosylphosphatidylinositol protein anchors of Trypanosoma cruzi Heise, Norton [UNIFESP] |
title_short |
Identification of complete precursors for the glycosylphosphatidylinositol protein anchors of Trypanosoma cruzi |
title_full |
Identification of complete precursors for the glycosylphosphatidylinositol protein anchors of Trypanosoma cruzi |
title_fullStr |
Identification of complete precursors for the glycosylphosphatidylinositol protein anchors of Trypanosoma cruzi |
title_full_unstemmed |
Identification of complete precursors for the glycosylphosphatidylinositol protein anchors of Trypanosoma cruzi |
title_sort |
Identification of complete precursors for the glycosylphosphatidylinositol protein anchors of Trypanosoma cruzi |
author |
Heise, Norton [UNIFESP] |
author_facet |
Heise, Norton [UNIFESP] Raper, J. Buxbaum, L. U. Peranovich, TMS Almeida, MLC |
author_role |
author |
author2 |
Raper, J. Buxbaum, L. U. Peranovich, TMS Almeida, MLC |
author2_role |
author author author author |
dc.contributor.institution.none.fl_str_mv |
Universidade Federal de São Paulo (UNIFESP) JOHNS HOPKINS UNIV |
dc.contributor.author.fl_str_mv |
Heise, Norton [UNIFESP] Raper, J. Buxbaum, L. U. Peranovich, TMS Almeida, MLC |
description |
The survival of Trypanosoma cruzi, the causative agent of Chagas' disease, depends vitally on proteins and glycoconjugates that mediate the parasite/host interaction. Since most of these molecules are attached to the membrane by glycosylphosphatidylinositol (GPI), alternative means of chemotherapeutic intervention might emerge from GPI biosynthesis studies. The structure of the major 1G7 antigen GPI has been fully characterized by us (Guther, M. L. S., Cardoso de Almeida, M. L., Yoshida, N., and Ferguson, M. A. J. (1992) J. Biol. Chem. 267, 6820-6828; Heise, N., Cardoso de Almeida, M. L., and Ferguson, M. A. J. (1995) Mel. Biochem. Parasitol. 70, 71-84), and based on its properties we now report the complete precursor glycolipids predicted to be transferred to the nascent protein. Migrating closely to Trypanosoma brucei glycolipid A on TLC, such species, named glycolipids A-like 1 and A-like 2, were labeled with tritiated palmitic acid, myo-inositol, glucosamine, and mannose, but surprisingly only the less polar glycolipid A-like 1 incorporated ethanolamine. The predicted products following nitrous acid deamination and digestion with phospholipases A(2), C, and D confirmed their GPI nature. Evidence that they may represent the anchor transferred to the 1G7 antigen came from the following analyses: (i) alpha-mannosidase treatments indicated that only one mannose was amenable to removal; (ii) their lipid moiety was identified as sn-l-alkyl-a-acylglycerol due to their sensitivity to phospholipase A(2) (PLA(2)), mild base and by direct high performance TLC analysis of the corresponding benzoylated diradylglycerol components; and (iii) both glycolipids incorporated H-3-fatty acid only in the sn-2- and not in the sn-l-alkyl position as previously found in the GPI of the mature 1G7 antigen. Based on the differential [H-3]ethanolamine incorporation pattern and the recent report that an aminoethylphosphonic acid (AEP) replaces ethanolamine phosphate (EtNH(2)-PO4) in the GPI in epimastigote sialoglycoproteins (Previato, J. O., Jones, C., Xavier, M. T., Wait, R., Travassos, L. R., Parodi, A. J., and Mendonca-Previato, L. (1995) J. Biol. Chem. 270, 7241-7250) it is proposed that glycolipid A-like 2 contains AEP and A-like 1 EtNH(2)-PO4. In the in vitro cell-free system both glycolipids were synthesized simultaneously and do not seem to bear a precursor/product relationship. Among the various components synthesized in vitro a glycolipid C-like corresponding to a form of glycolipid A-like 1 acylated on the inositol was also characterized. Phenylmethylsulfonyl fluoride, an inhibitor known to block the addition of ethanolamine phosphate in T. brucei but not in mammalian cells, also inhibits the synthesis of glycolipids A like and C-like in T. cruzi, indicating that the putative trypanosome EtNH(2)-PO4/AEP transferase(s) might represent a potential target for chemotherapy. |
publishDate |
1996 |
dc.date.issued.fl_str_mv |
1996-07-12 |
dc.date.accessioned.fl_str_mv |
2018-06-15T17:53:05Z |
dc.date.available.fl_str_mv |
2018-06-15T17:53:05Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.citation.fl_str_mv |
Journal Of Biological Chemistry. Bethesda: Amer Soc Biochemistry Molecular Biology Inc, v. 271, n. 28, p. 16877-16887, 1996. |
dc.identifier.uri.fl_str_mv |
http://repositorio.unifesp.br/11600/44205 http://doi.org/10.1074/jbc.271.28.16877 |
dc.identifier.issn.none.fl_str_mv |
0021-9258 |
dc.identifier.doi.none.fl_str_mv |
10.1074/jbc.271.28.16877 |
dc.identifier.wos.none.fl_str_mv |
WOS:A1996UX12600070 |
identifier_str_mv |
Journal Of Biological Chemistry. Bethesda: Amer Soc Biochemistry Molecular Biology Inc, v. 271, n. 28, p. 16877-16887, 1996. 0021-9258 10.1074/jbc.271.28.16877 WOS:A1996UX12600070 |
url |
http://repositorio.unifesp.br/11600/44205 http://doi.org/10.1074/jbc.271.28.16877 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.ispartof.none.fl_str_mv |
Journal Of Biological Chemistry |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
16877-16887 |
dc.publisher.none.fl_str_mv |
Amer Soc Biochemistry Molecular Biology Inc |
publisher.none.fl_str_mv |
Amer Soc Biochemistry Molecular Biology Inc |
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 |
|
_version_ |
1802764124555313152 |