Extracellular ATP triggers proteolysis and cytosolic Ca2+ rise in Plasmodium berghei and Plasmodium yoelii malaria parasites
Autor(a) principal: | |
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Data de Publicação: | 2012 |
Outros Autores: | , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNIFESP |
Texto Completo: | http://repositorio.unifesp.br/handle/11600/34713 http://dx.doi.org/10.1186/1475-2875-11-69 |
Resumo: | Background: Plasmodium has a complex cell biology and it is essential to dissect the cell-signalling pathways underlying its survival within the host.Methods: Using the fluorescence resonance energy transfer (FRET) peptide substrate Abz-AIKFFARQ-EDDnp and Fluo4/AM, the effects of extracellular ATP on triggering proteolysis and Ca2+ signalling in Plasmodium berghei and Plasmodium yoelii malaria parasites were investigated.Results: the protease activity was blocked in the presence of the purinergic receptor blockers suramin (50 mu M) and PPADS (50 mu M) or the extracellular and intracellular calcium chelators EGTA (5 mM) and BAPTA/AM (25, 100, 200 and 500 mu M), respectively for P. yoelii and P. berghei. Addition of ATP (50, 70, 200 and 250 mu M) to isolated parasites previously loaded with Fluo4/AM in a Ca2+-containing medium led to an increase in cytosolic calcium. This rise was blocked by pre-incubating the parasites with either purinergic antagonists PPADS (50 mu M), TNP-ATP (50 mu M) or the purinergic blockers KN-62 (10 mu M) and Ip5I (10 mu M). Incubating P. berghei infected cells with KN-62 (200 mu M) resulted in a changed profile of merozoite surface protein 1 (MSP1) processing as revealed by western blot assays. Moreover incubating P. berghei for 17 h with KN-62 (10 mu M) led to an increase in rings forms (82% +/- 4, n = 11) and a decrease in trophozoite forms (18% +/- 4, n = 11).Conclusions: the data clearly show that purinergic signalling modulates P. berghei protease(s) activity and that MSP1 is one target in this pathway. |
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Cruz, Laura NogueiraJuliano, Maria Aparecida [UNIFESP]Budu, AlexandreJuliano, Luiz [UNIFESP]Holder, Anthony A.Blackman, Michael J.Garcia, Celia R. S.Universidade de São Paulo (USP)Universidade Federal de São Paulo (UNIFESP)MRC Natl Inst Med Res2016-01-24T14:26:58Z2016-01-24T14:26:58Z2012-03-15Malaria Journal. London: Biomed Central Ltd, v. 11, 11 p., 2012.1475-2875http://repositorio.unifesp.br/handle/11600/34713http://dx.doi.org/10.1186/1475-2875-11-69WOS000304544700001.pdf10.1186/1475-2875-11-69WOS:000304544700001Background: Plasmodium has a complex cell biology and it is essential to dissect the cell-signalling pathways underlying its survival within the host.Methods: Using the fluorescence resonance energy transfer (FRET) peptide substrate Abz-AIKFFARQ-EDDnp and Fluo4/AM, the effects of extracellular ATP on triggering proteolysis and Ca2+ signalling in Plasmodium berghei and Plasmodium yoelii malaria parasites were investigated.Results: the protease activity was blocked in the presence of the purinergic receptor blockers suramin (50 mu M) and PPADS (50 mu M) or the extracellular and intracellular calcium chelators EGTA (5 mM) and BAPTA/AM (25, 100, 200 and 500 mu M), respectively for P. yoelii and P. berghei. Addition of ATP (50, 70, 200 and 250 mu M) to isolated parasites previously loaded with Fluo4/AM in a Ca2+-containing medium led to an increase in cytosolic calcium. This rise was blocked by pre-incubating the parasites with either purinergic antagonists PPADS (50 mu M), TNP-ATP (50 mu M) or the purinergic blockers KN-62 (10 mu M) and Ip5I (10 mu M). Incubating P. berghei infected cells with KN-62 (200 mu M) resulted in a changed profile of merozoite surface protein 1 (MSP1) processing as revealed by western blot assays. Moreover incubating P. berghei for 17 h with KN-62 (10 mu M) led to an increase in rings forms (82% +/- 4, n = 11) and a decrease in trophozoite forms (18% +/- 4, n = 11).Conclusions: the data clearly show that purinergic signalling modulates P. berghei protease(s) activity and that MSP1 is one target in this pathway.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)UK Medical Research CouncilEU through the Network of Excellence EviMalaRUniv São Paulo, Dept Physiol, Inst Biociencias, BR-05508900 São Paulo, SP, BrazilUniversidade Federal de São Paulo, Escola Paulista Med, Dept Biophys, BR-04044020 São Paulo, SP, BrazilMRC Natl Inst Med Res, Div Parasitol, London NW7 1AA, EnglandUniv São Paulo, Inst Ciencias Biomed, Dept Parasitol, BR-05508900 São Paulo, SP, BrazilUniversidade Federal de São Paulo, Escola Paulista Med, Dept Biophys, BR-04044020 São Paulo, SP, BrazilUK Medical Research Council: U117532067UK Medical Research Council: U117532063EU through the Network of Excellence EviMalaR: Health-2009-2.3.2-1-242095Web of Science11engBiomed Central LtdMalaria JournalATPPurinergic receptorMalariaPlasmodium bergheiPlasmodium yoeliiProtease activityCalcium modulationMerozoite surface protein 1Extracellular ATP triggers proteolysis and cytosolic Ca2+ rise in Plasmodium berghei and Plasmodium yoelii malaria parasitesinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UNIFESPinstname:Universidade Federal de São Paulo (UNIFESP)instacron:UNIFESPORIGINALWOS000304544700001.pdfapplication/pdf710046${dspace.ui.url}/bitstream/11600/34713/1/WOS000304544700001.pdf1a5647aace7400ee682637ef1b364a16MD51open accessTEXTWOS000304544700001.pdf.txtWOS000304544700001.pdf.txtExtracted texttext/plain48945${dspace.ui.url}/bitstream/11600/34713/2/WOS000304544700001.pdf.txtef6b7b93058732b04b2ca43d9ede3fefMD52open access11600/347132023-01-12 21:52:30.486open accessoai:repositorio.unifesp.br:11600/34713Repositório InstitucionalPUBhttp://www.repositorio.unifesp.br/oai/requestopendoar:34652023-05-25T12:11:09.638276Repositório Institucional da UNIFESP - Universidade Federal de São Paulo (UNIFESP)false |
dc.title.en.fl_str_mv |
Extracellular ATP triggers proteolysis and cytosolic Ca2+ rise in Plasmodium berghei and Plasmodium yoelii malaria parasites |
title |
Extracellular ATP triggers proteolysis and cytosolic Ca2+ rise in Plasmodium berghei and Plasmodium yoelii malaria parasites |
spellingShingle |
Extracellular ATP triggers proteolysis and cytosolic Ca2+ rise in Plasmodium berghei and Plasmodium yoelii malaria parasites Cruz, Laura Nogueira ATP Purinergic receptor Malaria Plasmodium berghei Plasmodium yoelii Protease activity Calcium modulation Merozoite surface protein 1 |
title_short |
Extracellular ATP triggers proteolysis and cytosolic Ca2+ rise in Plasmodium berghei and Plasmodium yoelii malaria parasites |
title_full |
Extracellular ATP triggers proteolysis and cytosolic Ca2+ rise in Plasmodium berghei and Plasmodium yoelii malaria parasites |
title_fullStr |
Extracellular ATP triggers proteolysis and cytosolic Ca2+ rise in Plasmodium berghei and Plasmodium yoelii malaria parasites |
title_full_unstemmed |
Extracellular ATP triggers proteolysis and cytosolic Ca2+ rise in Plasmodium berghei and Plasmodium yoelii malaria parasites |
title_sort |
Extracellular ATP triggers proteolysis and cytosolic Ca2+ rise in Plasmodium berghei and Plasmodium yoelii malaria parasites |
author |
Cruz, Laura Nogueira |
author_facet |
Cruz, Laura Nogueira Juliano, Maria Aparecida [UNIFESP] Budu, Alexandre Juliano, Luiz [UNIFESP] Holder, Anthony A. Blackman, Michael J. Garcia, Celia R. S. |
author_role |
author |
author2 |
Juliano, Maria Aparecida [UNIFESP] Budu, Alexandre Juliano, Luiz [UNIFESP] Holder, Anthony A. Blackman, Michael J. Garcia, Celia R. S. |
author2_role |
author author author author author author |
dc.contributor.institution.none.fl_str_mv |
Universidade de São Paulo (USP) Universidade Federal de São Paulo (UNIFESP) MRC Natl Inst Med Res |
dc.contributor.author.fl_str_mv |
Cruz, Laura Nogueira Juliano, Maria Aparecida [UNIFESP] Budu, Alexandre Juliano, Luiz [UNIFESP] Holder, Anthony A. Blackman, Michael J. Garcia, Celia R. S. |
dc.subject.eng.fl_str_mv |
ATP Purinergic receptor Malaria Plasmodium berghei Plasmodium yoelii Protease activity Calcium modulation Merozoite surface protein 1 |
topic |
ATP Purinergic receptor Malaria Plasmodium berghei Plasmodium yoelii Protease activity Calcium modulation Merozoite surface protein 1 |
description |
Background: Plasmodium has a complex cell biology and it is essential to dissect the cell-signalling pathways underlying its survival within the host.Methods: Using the fluorescence resonance energy transfer (FRET) peptide substrate Abz-AIKFFARQ-EDDnp and Fluo4/AM, the effects of extracellular ATP on triggering proteolysis and Ca2+ signalling in Plasmodium berghei and Plasmodium yoelii malaria parasites were investigated.Results: the protease activity was blocked in the presence of the purinergic receptor blockers suramin (50 mu M) and PPADS (50 mu M) or the extracellular and intracellular calcium chelators EGTA (5 mM) and BAPTA/AM (25, 100, 200 and 500 mu M), respectively for P. yoelii and P. berghei. Addition of ATP (50, 70, 200 and 250 mu M) to isolated parasites previously loaded with Fluo4/AM in a Ca2+-containing medium led to an increase in cytosolic calcium. This rise was blocked by pre-incubating the parasites with either purinergic antagonists PPADS (50 mu M), TNP-ATP (50 mu M) or the purinergic blockers KN-62 (10 mu M) and Ip5I (10 mu M). Incubating P. berghei infected cells with KN-62 (200 mu M) resulted in a changed profile of merozoite surface protein 1 (MSP1) processing as revealed by western blot assays. Moreover incubating P. berghei for 17 h with KN-62 (10 mu M) led to an increase in rings forms (82% +/- 4, n = 11) and a decrease in trophozoite forms (18% +/- 4, n = 11).Conclusions: the data clearly show that purinergic signalling modulates P. berghei protease(s) activity and that MSP1 is one target in this pathway. |
publishDate |
2012 |
dc.date.issued.fl_str_mv |
2012-03-15 |
dc.date.accessioned.fl_str_mv |
2016-01-24T14:26:58Z |
dc.date.available.fl_str_mv |
2016-01-24T14:26:58Z |
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 |
Malaria Journal. London: Biomed Central Ltd, v. 11, 11 p., 2012. |
dc.identifier.uri.fl_str_mv |
http://repositorio.unifesp.br/handle/11600/34713 http://dx.doi.org/10.1186/1475-2875-11-69 |
dc.identifier.issn.none.fl_str_mv |
1475-2875 |
dc.identifier.file.none.fl_str_mv |
WOS000304544700001.pdf |
dc.identifier.doi.none.fl_str_mv |
10.1186/1475-2875-11-69 |
dc.identifier.wos.none.fl_str_mv |
WOS:000304544700001 |
identifier_str_mv |
Malaria Journal. London: Biomed Central Ltd, v. 11, 11 p., 2012. 1475-2875 WOS000304544700001.pdf 10.1186/1475-2875-11-69 WOS:000304544700001 |
url |
http://repositorio.unifesp.br/handle/11600/34713 http://dx.doi.org/10.1186/1475-2875-11-69 |
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eng |
language |
eng |
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Malaria Journal |
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11 |
dc.publisher.none.fl_str_mv |
Biomed Central Ltd |
publisher.none.fl_str_mv |
Biomed Central Ltd |
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