Calcium uptake and proton transport by acidocalcisomes of toxoplasma gondii
Autor(a) principal: | |
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Data de Publicação: | 2011 |
Outros Autores: | , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional do INMETRO |
Texto Completo: | http://hdl.handle.net/10926/1936 |
Resumo: | Acidocalcisomes are acidic calcium stores found in diverse organisms, being conserved from bacteria to humans. They possess an acidic matrix that contains several cations bound to phosphates, which are mainly present in the form of short and long polyphosphate chains. Their matrix is acidified through the action of proton pumps such as a vacuolar proton ATPase and a vacuolar proton pyrophosphatase. Calcium uptake occurs through a Ca2+/H+ countertransporting ATPase located in the membrane of the organelle. Acidocalcisomes have been identified in a variety of microorganisms, including Apicomplexan parasites such as Plasmodium and Eimeria species, and in Toxoplasma gondii. We report the purification and characterization of an acidocalcisome fraction from T. gondii tachyzoites after subcellular fractionation and further discontinuous iodixanol gradient purification. Proton and calcium transport activities in the fraction were characterized by fluorescence microscopy and spectrophotometric methods using acridine orange and arsenazo III, respectively. This work will facilitate the understanding of the function of acidocalcisomes in Apicomplexan parasites, as we can now isolate highly purified fractions that could be used for proteomic analysis to find proteins that may clarify the biogenesis of these organelles. |
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info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleCalcium uptake and proton transport by acidocalcisomes of toxoplasma gondii20112013-01-16T14:37:41Z2013-01-16T14:37:41ZAcidocalcisomes are acidic calcium stores found in diverse organisms, being conserved from bacteria to humans. They possess an acidic matrix that contains several cations bound to phosphates, which are mainly present in the form of short and long polyphosphate chains. Their matrix is acidified through the action of proton pumps such as a vacuolar proton ATPase and a vacuolar proton pyrophosphatase. Calcium uptake occurs through a Ca2+/H+ countertransporting ATPase located in the membrane of the organelle. Acidocalcisomes have been identified in a variety of microorganisms, including Apicomplexan parasites such as Plasmodium and Eimeria species, and in Toxoplasma gondii. We report the purification and characterization of an acidocalcisome fraction from T. gondii tachyzoites after subcellular fractionation and further discontinuous iodixanol gradient purification. Proton and calcium transport activities in the fraction were characterized by fluorescence microscopy and spectrophotometric methods using acridine orange and arsenazo III, respectively. This work will facilitate the understanding of the function of acidocalcisomes in Apicomplexan parasites, as we can now isolate highly purified fractions that could be used for proteomic analysis to find proteins that may clarify the biogenesis of these organelles.9 p. : il., tab.Submitted by Lívia Oliveira (livia.oliveira2@hotmail.com) on 2012-07-26T14:45:09Z No. of bitstreams: 1 Miranda4_2011.pdf: 615111 bytes, checksum: 48c7eb155512fcccc85637b8ff1b9f97 (MD5)Approved for entry into archive by Regina Mello(mrmello@inmetro.gov.br) on 2013-01-16T14:37:41Z (GMT) No. of bitstreams: 1 Miranda4_2011.pdf: 615111 bytes, checksum: 48c7eb155512fcccc85637b8ff1b9f97 (MD5)Made available in DSpace on 2013-01-16T14:37:41Z (GMT). No. of bitstreams: 1 Miranda4_2011.pdf: 615111 bytes, checksum: 48c7eb155512fcccc85637b8ff1b9f97 (MD5) Previous issue date: 2011enghttp://hdl.handle.net/10926/1936DMD_hdl_10926/1936ROHLOFF, Peter et al. Calcium uptake and proton transport by acidocalcisomes of toxoplasma gondii. Plos One, v. 6, n. 4, 2011.Rohloff, PeterMiranda, Kildare Rocha deRodrigues, Juliany Cola FernandesFang, JianminGalizzi, MelinaPlattner, HelmutHentschel, JoachimMoreno, Silvia Noemi Josefinainfo:eu-repo/semantics/openAccessreponame:Repositório Institucional do INMETROinstname:Instituto Nacional de Metrologia, Qualidade e Tecnologia (INMETRO)instacron:INMETRORohloff_2011.pdfhttp://xrepo01s.inmetro.gov.br/bitstream/10926/1936/1/Rohloff_2011.pdfapplication/pdf615111http://xrepo01s.inmetro.gov.br/bitstream/10926/1936/1/Rohloff_2011.pdf48c7eb155512fcccc85637b8ff1b9f97MD510926_1936_1Rohloff_2011.pdf.txthttp://xrepo01s.inmetro.gov.br/bitstream/10926/1936/6/Rohloff_2011.pdf.txttext/plain43818http://xrepo01s.inmetro.gov.br/bitstream/10926/1936/6/Rohloff_2011.pdf.txt12c7087a259c742552e516242cadd51aMD510926_1936_62024-06-10T15:24:56Zoai:xrepo01s.inmetro.gov.br:10926/1936Repositório de Publicaçõeshttp://repositorios.inmetro.gov.br/oai/requestopendoar:2013-10-02T13:37:46Repositório Institucional do INMETRO - Instituto Nacional de Metrologia, Qualidade e Tecnologia (INMETRO)false |
dc.title.none.fl_str_mv |
Calcium uptake and proton transport by acidocalcisomes of toxoplasma gondii |
title |
Calcium uptake and proton transport by acidocalcisomes of toxoplasma gondii |
spellingShingle |
Calcium uptake and proton transport by acidocalcisomes of toxoplasma gondii Rohloff, Peter |
title_short |
Calcium uptake and proton transport by acidocalcisomes of toxoplasma gondii |
title_full |
Calcium uptake and proton transport by acidocalcisomes of toxoplasma gondii |
title_fullStr |
Calcium uptake and proton transport by acidocalcisomes of toxoplasma gondii |
title_full_unstemmed |
Calcium uptake and proton transport by acidocalcisomes of toxoplasma gondii |
title_sort |
Calcium uptake and proton transport by acidocalcisomes of toxoplasma gondii |
author |
Rohloff, Peter |
author_facet |
Rohloff, Peter Miranda, Kildare Rocha de Rodrigues, Juliany Cola Fernandes Fang, Jianmin Galizzi, Melina Plattner, Helmut Hentschel, Joachim Moreno, Silvia Noemi Josefina |
author_role |
author |
author2 |
Miranda, Kildare Rocha de Rodrigues, Juliany Cola Fernandes Fang, Jianmin Galizzi, Melina Plattner, Helmut Hentschel, Joachim Moreno, Silvia Noemi Josefina |
author2_role |
author author author author author author author |
dc.contributor.author.fl_str_mv |
Rohloff, Peter Miranda, Kildare Rocha de Rodrigues, Juliany Cola Fernandes Fang, Jianmin Galizzi, Melina Plattner, Helmut Hentschel, Joachim Moreno, Silvia Noemi Josefina |
description |
Acidocalcisomes are acidic calcium stores found in diverse organisms, being conserved from bacteria to humans. They possess an acidic matrix that contains several cations bound to phosphates, which are mainly present in the form of short and long polyphosphate chains. Their matrix is acidified through the action of proton pumps such as a vacuolar proton ATPase and a vacuolar proton pyrophosphatase. Calcium uptake occurs through a Ca2+/H+ countertransporting ATPase located in the membrane of the organelle. Acidocalcisomes have been identified in a variety of microorganisms, including Apicomplexan parasites such as Plasmodium and Eimeria species, and in Toxoplasma gondii. We report the purification and characterization of an acidocalcisome fraction from T. gondii tachyzoites after subcellular fractionation and further discontinuous iodixanol gradient purification. Proton and calcium transport activities in the fraction were characterized by fluorescence microscopy and spectrophotometric methods using acridine orange and arsenazo III, respectively. This work will facilitate the understanding of the function of acidocalcisomes in Apicomplexan parasites, as we can now isolate highly purified fractions that could be used for proteomic analysis to find proteins that may clarify the biogenesis of these organelles. |
publishDate |
2011 |
dc.date.issued.fl_str_mv |
2011 |
dc.date.available.fl_str_mv |
2013-01-16T14:37:41Z |
dc.date.accessioned.fl_str_mv |
2013-01-16T14:37:41Z |
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.uri.fl_str_mv |
http://hdl.handle.net/10926/1936 DMD_hdl_10926/1936 |
dc.identifier.citation.fl_str_mv |
ROHLOFF, Peter et al. Calcium uptake and proton transport by acidocalcisomes of toxoplasma gondii. Plos One, v. 6, n. 4, 2011. |
url |
http://hdl.handle.net/10926/1936 |
identifier_str_mv |
DMD_hdl_10926/1936 ROHLOFF, Peter et al. Calcium uptake and proton transport by acidocalcisomes of toxoplasma gondii. Plos One, v. 6, n. 4, 2011. |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.bitstream.fl_str_mv |
application/pdf text/plain |
dc.source.none.fl_str_mv |
reponame:Repositório Institucional do INMETRO instname:Instituto Nacional de Metrologia, Qualidade e Tecnologia (INMETRO) instacron:INMETRO |
instname_str |
Instituto Nacional de Metrologia, Qualidade e Tecnologia (INMETRO) |
instacron_str |
INMETRO |
institution |
INMETRO |
reponame_str |
Repositório Institucional do INMETRO |
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Repositório Institucional do INMETRO |
repository.name.fl_str_mv |
Repositório Institucional do INMETRO - Instituto Nacional de Metrologia, Qualidade e Tecnologia (INMETRO) |
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