Dihydroorotate dehydrogenase of Toxoplasma gondii - kinetic characterization and intracellular localization
The pyrimidine biosynthesis patway in the protozoan pathogen Toxoplosma gondii is essenial for parasite growth during infection. To investigate the properties of Dihydroorotate dehydrogenase (TgDHOD. TgDHOD exhibited a specific activity of 83.8 U/mg, a kcat of 89.2 sec-1 ± 1.5. a Km = 60.3 ±0.002 æM...
- Autores:
-
Hortua Triana, Miryam Andrea
- Tipo de recurso:
- Doctoral thesis
- Fecha de publicación:
- 2010
- Institución:
- Universidad de los Andes
- Repositorio:
- Séneca: repositorio Uniandes
- Idioma:
- eng
- OAI Identifier:
- oai:repositorio.uniandes.edu.co:1992/7764
- Acceso en línea:
- http://hdl.handle.net/1992/7764
- Palabra clave:
- Dihydroorotate dehydrogenase - Investigaciones
Toxoplasma gondii - Investigaciones
Pirimidinas - Biosíntesis - Investigaciones
Plástidos - Investigaciones
Biología
- Rights
- openAccess
- License
- http://creativecommons.org/licenses/by-nc-sa/4.0/
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Al consultar y hacer uso de este recurso, está aceptando las condiciones de uso establecidas por los autores.http://creativecommons.org/licenses/by-nc-sa/4.0/info:eu-repo/semantics/openAccesshttp://purl.org/coar/access_right/c_abf2Zimmermann, Barbara Hanna81271423-588b-479b-a844-0d7443608a49500Hortua Triana, Miryam Andrea1312e14c-74bd-4d94-a33f-df2f024c80765002018-09-27T16:38:01Z2018-09-27T16:38:01Z2010http://hdl.handle.net/1992/776410.57784/1992/7764u429388.pdfinstname:Universidad de los Andesreponame:Repositorio Institucional Sénecarepourl:https://repositorio.uniandes.edu.co/The pyrimidine biosynthesis patway in the protozoan pathogen Toxoplosma gondii is essenial for parasite growth during infection. To investigate the properties of Dihydroorotate dehydrogenase (TgDHOD. TgDHOD exhibited a specific activity of 83.8 U/mg, a kcat of 89.2 sec-1 ± 1.5. a Km = 60.3 ±0.002 æM for L-dihydroorotate, and a Km = 28.9 æM ± 1.8 for decylubiquinone (QD). Quinones lacking or having short isoprenoid side chains yielded lower kcats and higher Kms than QD. As expected, fumarate was a poor electron acceptor for this family 2 DHOD. The determined for Redoxal and A77-1726 were 253.3 æM ± 13.3 and 91.2 æM ±2.2, respectively. The enzyme was not significantly affected by brequinar or TTFA, known inhibitors of human DHOD, or by atovaquone. TgDHOD exhibits a 157 ? residue N-terminal extension, consistent with potential organellar targeting, but bioinformatic analysis failed to reveal a consensus subcellular destination, Preliminary inmunolocalization studies of TgDHOD performed in intra- and extracellular parasites treated with the polyclonal antibodies raised against purified recombinant TgDHOD exhibited fluorescence that appeared to colocalize with the apicoplast in fluorescence was observed in -10% of mitochondria stained with Mitotracker Red in extracellular parasites. These findings suggest that TgDHOD is associated with both the apicoplast and mitochondrion, making it a member of a growing list of a metabolic enzymes that are dual targeted in T. gondiiDoctor en Ciencias - BiologíaDoctorado83 hojasapplication/pdfengUniandesDoctorado en Ciencias - BiologíaFacultad de CienciasDepartamento de Ciencias Biológicasinstname:Universidad de los Andesreponame:Repositorio Institucional SénecaDihydroorotate dehydrogenase of Toxoplasma gondii - kinetic characterization and intracellular localizationTrabajo de grado - Doctoradoinfo:eu-repo/semantics/doctoralThesishttp://purl.org/coar/resource_type/c_db06http://purl.org/coar/version/c_970fb48d4fbd8a85Texthttp://purl.org/redcol/resource_type/TDDihydroorotate dehydrogenase - InvestigacionesToxoplasma gondii - InvestigacionesPirimidinas - Biosíntesis - InvestigacionesPlástidos - InvestigacionesBiologíaPublicationTEXTu429388.pdf.txtu429388.pdf.txtExtracted texttext/plain120399https://repositorio.uniandes.edu.co/bitstreams/84b4e81c-f873-48de-a68c-f00bb5cdce6f/download9eca11378a98daf13debf42d58fd5bdaMD56ORIGINALu429388.pdfapplication/pdf1916469https://repositorio.uniandes.edu.co/bitstreams/67daf557-88a7-4484-a049-af45e8ebca52/downloadb58552de5af2d986f444e6d99d1b14ccMD51THUMBNAILu429388.pdf.jpgu429388.pdf.jpgIM Thumbnailimage/jpeg6603https://repositorio.uniandes.edu.co/bitstreams/9a531e8e-bc4c-419d-b952-950a1c1af206/download4ab26ecc1a48ea390492dc797faf95e8MD571992/7764oai:repositorio.uniandes.edu.co:1992/77642024-11-14 10:33:14.289http://creativecommons.org/licenses/by-nc-sa/4.0/open.accesshttps://repositorio.uniandes.edu.coRepositorio institucional Sénecaadminrepositorio@uniandes.edu.co |
dc.title.es_CO.fl_str_mv |
Dihydroorotate dehydrogenase of Toxoplasma gondii - kinetic characterization and intracellular localization |
title |
Dihydroorotate dehydrogenase of Toxoplasma gondii - kinetic characterization and intracellular localization |
spellingShingle |
Dihydroorotate dehydrogenase of Toxoplasma gondii - kinetic characterization and intracellular localization Dihydroorotate dehydrogenase - Investigaciones Toxoplasma gondii - Investigaciones Pirimidinas - Biosíntesis - Investigaciones Plástidos - Investigaciones Biología |
title_short |
Dihydroorotate dehydrogenase of Toxoplasma gondii - kinetic characterization and intracellular localization |
title_full |
Dihydroorotate dehydrogenase of Toxoplasma gondii - kinetic characterization and intracellular localization |
title_fullStr |
Dihydroorotate dehydrogenase of Toxoplasma gondii - kinetic characterization and intracellular localization |
title_full_unstemmed |
Dihydroorotate dehydrogenase of Toxoplasma gondii - kinetic characterization and intracellular localization |
title_sort |
Dihydroorotate dehydrogenase of Toxoplasma gondii - kinetic characterization and intracellular localization |
dc.creator.fl_str_mv |
Hortua Triana, Miryam Andrea |
dc.contributor.advisor.none.fl_str_mv |
Zimmermann, Barbara Hanna |
dc.contributor.author.none.fl_str_mv |
Hortua Triana, Miryam Andrea |
dc.subject.keyword.es_CO.fl_str_mv |
Dihydroorotate dehydrogenase - Investigaciones Toxoplasma gondii - Investigaciones Pirimidinas - Biosíntesis - Investigaciones Plástidos - Investigaciones |
topic |
Dihydroorotate dehydrogenase - Investigaciones Toxoplasma gondii - Investigaciones Pirimidinas - Biosíntesis - Investigaciones Plástidos - Investigaciones Biología |
dc.subject.themes.none.fl_str_mv |
Biología |
description |
The pyrimidine biosynthesis patway in the protozoan pathogen Toxoplosma gondii is essenial for parasite growth during infection. To investigate the properties of Dihydroorotate dehydrogenase (TgDHOD. TgDHOD exhibited a specific activity of 83.8 U/mg, a kcat of 89.2 sec-1 ± 1.5. a Km = 60.3 ±0.002 æM for L-dihydroorotate, and a Km = 28.9 æM ± 1.8 for decylubiquinone (QD). Quinones lacking or having short isoprenoid side chains yielded lower kcats and higher Kms than QD. As expected, fumarate was a poor electron acceptor for this family 2 DHOD. The determined for Redoxal and A77-1726 were 253.3 æM ± 13.3 and 91.2 æM ±2.2, respectively. The enzyme was not significantly affected by brequinar or TTFA, known inhibitors of human DHOD, or by atovaquone. TgDHOD exhibits a 157 ? residue N-terminal extension, consistent with potential organellar targeting, but bioinformatic analysis failed to reveal a consensus subcellular destination, Preliminary inmunolocalization studies of TgDHOD performed in intra- and extracellular parasites treated with the polyclonal antibodies raised against purified recombinant TgDHOD exhibited fluorescence that appeared to colocalize with the apicoplast in fluorescence was observed in -10% of mitochondria stained with Mitotracker Red in extracellular parasites. These findings suggest that TgDHOD is associated with both the apicoplast and mitochondrion, making it a member of a growing list of a metabolic enzymes that are dual targeted in T. gondii |
publishDate |
2010 |
dc.date.issued.none.fl_str_mv |
2010 |
dc.date.accessioned.none.fl_str_mv |
2018-09-27T16:38:01Z |
dc.date.available.none.fl_str_mv |
2018-09-27T16:38:01Z |
dc.type.spa.fl_str_mv |
Trabajo de grado - Doctorado |
dc.type.coarversion.fl_str_mv |
http://purl.org/coar/version/c_970fb48d4fbd8a85 |
dc.type.driver.spa.fl_str_mv |
info:eu-repo/semantics/doctoralThesis |
dc.type.coar.spa.fl_str_mv |
http://purl.org/coar/resource_type/c_db06 |
dc.type.content.spa.fl_str_mv |
Text |
dc.type.redcol.spa.fl_str_mv |
http://purl.org/redcol/resource_type/TD |
format |
http://purl.org/coar/resource_type/c_db06 |
dc.identifier.uri.none.fl_str_mv |
http://hdl.handle.net/1992/7764 |
dc.identifier.doi.none.fl_str_mv |
10.57784/1992/7764 |
dc.identifier.pdf.none.fl_str_mv |
u429388.pdf |
dc.identifier.instname.spa.fl_str_mv |
instname:Universidad de los Andes |
dc.identifier.reponame.spa.fl_str_mv |
reponame:Repositorio Institucional Séneca |
dc.identifier.repourl.spa.fl_str_mv |
repourl:https://repositorio.uniandes.edu.co/ |
url |
http://hdl.handle.net/1992/7764 |
identifier_str_mv |
10.57784/1992/7764 u429388.pdf instname:Universidad de los Andes reponame:Repositorio Institucional Séneca repourl:https://repositorio.uniandes.edu.co/ |
dc.language.iso.es_CO.fl_str_mv |
eng |
language |
eng |
dc.rights.uri.*.fl_str_mv |
http://creativecommons.org/licenses/by-nc-sa/4.0/ |
dc.rights.accessrights.spa.fl_str_mv |
info:eu-repo/semantics/openAccess |
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http://purl.org/coar/access_right/c_abf2 |
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http://creativecommons.org/licenses/by-nc-sa/4.0/ http://purl.org/coar/access_right/c_abf2 |
eu_rights_str_mv |
openAccess |
dc.format.extent.es_CO.fl_str_mv |
83 hojas |
dc.format.mimetype.es_CO.fl_str_mv |
application/pdf |
dc.publisher.es_CO.fl_str_mv |
Uniandes |
dc.publisher.program.es_CO.fl_str_mv |
Doctorado en Ciencias - Biología |
dc.publisher.faculty.es_CO.fl_str_mv |
Facultad de Ciencias |
dc.publisher.department.spa.fl_str_mv |
Departamento de Ciencias Biológicas |
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Universidad de los Andes |
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Repositorio Institucional Séneca |
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Repositorio Institucional Séneca |
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