Teaching about citric acid cycle using plant mitochondrial preparations: Some assays for use in laboratory courses
Autor(a) principal: | |
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Data de Publicação: | 2005 |
Outros Autores: | , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | http://hdl.handle.net/10316/8146 https://doi.org/10.1002/bmb.2005.494033022444 |
Resumo: | Potato tubers and turnip roots were used to prepare purified mitochondria for laboratory practical work in the teaching of the citric acid cycle (TCA cycle). Plant mitochondria are particularly advantageous over the animal fractions to demonstrate the TCA cycle enzymatic steps, by using simple techniques to measure O2 consumption and transmembrane potential (ΔΨ). The several TCA cycle intermediates induce specific enzyme activities, which can be identified by respiratory parameters. Such a strategy is also used to evidence properties of the TCA cycle enzymes: ADP stimulation of isocitrate dehydrogenase and α-ketoglutarate dehydrogenase; activation by citrate of downstream oxidation steps, e.g. succinate dehydrogenase; and regulation of the activity of isocitrate dehydrogenase by citrate action on the citrate/isocitrate carrier. Furthermore, it has been demonstrated that, in the absence of exogenous Mg2+, isocitrate-dependent respiration favors the alternative oxidase pathway, as judged by changes of the ADP/O elicited by the inhibitor n-propyl galate. These are some examples of assays related with TCA cycle intermediates we can use in laboratory courses. |
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Teaching about citric acid cycle using plant mitochondrial preparations: Some assays for use in laboratory coursesPotato tubers and turnip roots were used to prepare purified mitochondria for laboratory practical work in the teaching of the citric acid cycle (TCA cycle). Plant mitochondria are particularly advantageous over the animal fractions to demonstrate the TCA cycle enzymatic steps, by using simple techniques to measure O2 consumption and transmembrane potential (ΔΨ). The several TCA cycle intermediates induce specific enzyme activities, which can be identified by respiratory parameters. Such a strategy is also used to evidence properties of the TCA cycle enzymes: ADP stimulation of isocitrate dehydrogenase and α-ketoglutarate dehydrogenase; activation by citrate of downstream oxidation steps, e.g. succinate dehydrogenase; and regulation of the activity of isocitrate dehydrogenase by citrate action on the citrate/isocitrate carrier. Furthermore, it has been demonstrated that, in the absence of exogenous Mg2+, isocitrate-dependent respiration favors the alternative oxidase pathway, as judged by changes of the ADP/O elicited by the inhibitor n-propyl galate. These are some examples of assays related with TCA cycle intermediates we can use in laboratory courses.2005info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articlehttp://hdl.handle.net/10316/8146http://hdl.handle.net/10316/8146https://doi.org/10.1002/bmb.2005.494033022444engBiochemistry and Molecular Biology Education. 33:2 (2005) 128-132Vicente, Joaquim A. F.Gomes-Santos, Carina S. S.Sousa, Ana Paula M.Madeira, Vítor M. C.info:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2021-11-02T09:49:35Zoai:estudogeral.uc.pt:10316/8146Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T20:55:51.759342Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse |
dc.title.none.fl_str_mv |
Teaching about citric acid cycle using plant mitochondrial preparations: Some assays for use in laboratory courses |
title |
Teaching about citric acid cycle using plant mitochondrial preparations: Some assays for use in laboratory courses |
spellingShingle |
Teaching about citric acid cycle using plant mitochondrial preparations: Some assays for use in laboratory courses Vicente, Joaquim A. F. |
title_short |
Teaching about citric acid cycle using plant mitochondrial preparations: Some assays for use in laboratory courses |
title_full |
Teaching about citric acid cycle using plant mitochondrial preparations: Some assays for use in laboratory courses |
title_fullStr |
Teaching about citric acid cycle using plant mitochondrial preparations: Some assays for use in laboratory courses |
title_full_unstemmed |
Teaching about citric acid cycle using plant mitochondrial preparations: Some assays for use in laboratory courses |
title_sort |
Teaching about citric acid cycle using plant mitochondrial preparations: Some assays for use in laboratory courses |
author |
Vicente, Joaquim A. F. |
author_facet |
Vicente, Joaquim A. F. Gomes-Santos, Carina S. S. Sousa, Ana Paula M. Madeira, Vítor M. C. |
author_role |
author |
author2 |
Gomes-Santos, Carina S. S. Sousa, Ana Paula M. Madeira, Vítor M. C. |
author2_role |
author author author |
dc.contributor.author.fl_str_mv |
Vicente, Joaquim A. F. Gomes-Santos, Carina S. S. Sousa, Ana Paula M. Madeira, Vítor M. C. |
description |
Potato tubers and turnip roots were used to prepare purified mitochondria for laboratory practical work in the teaching of the citric acid cycle (TCA cycle). Plant mitochondria are particularly advantageous over the animal fractions to demonstrate the TCA cycle enzymatic steps, by using simple techniques to measure O2 consumption and transmembrane potential (ΔΨ). The several TCA cycle intermediates induce specific enzyme activities, which can be identified by respiratory parameters. Such a strategy is also used to evidence properties of the TCA cycle enzymes: ADP stimulation of isocitrate dehydrogenase and α-ketoglutarate dehydrogenase; activation by citrate of downstream oxidation steps, e.g. succinate dehydrogenase; and regulation of the activity of isocitrate dehydrogenase by citrate action on the citrate/isocitrate carrier. Furthermore, it has been demonstrated that, in the absence of exogenous Mg2+, isocitrate-dependent respiration favors the alternative oxidase pathway, as judged by changes of the ADP/O elicited by the inhibitor n-propyl galate. These are some examples of assays related with TCA cycle intermediates we can use in laboratory courses. |
publishDate |
2005 |
dc.date.none.fl_str_mv |
2005 |
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/10316/8146 http://hdl.handle.net/10316/8146 https://doi.org/10.1002/bmb.2005.494033022444 |
url |
http://hdl.handle.net/10316/8146 https://doi.org/10.1002/bmb.2005.494033022444 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Biochemistry and Molecular Biology Education. 33:2 (2005) 128-132 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.source.none.fl_str_mv |
reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação instacron:RCAAP |
instname_str |
Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
instacron_str |
RCAAP |
institution |
RCAAP |
reponame_str |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
collection |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
repository.name.fl_str_mv |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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