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...

Full description

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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spelling 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
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dc.type.driver.spa.fl_str_mv info:eu-repo/semantics/doctoralThesis
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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
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url http://hdl.handle.net/1992/7764
identifier_str_mv 10.57784/1992/7764
u429388.pdf
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reponame:Repositorio Institucional Séneca
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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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rights_invalid_str_mv 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
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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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reponame:Repositorio Institucional Séneca
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