Clonagem e caracterização molecular da fosforribosil pirofosfato sintase (PRPP sintase) de cana-de-açucar.
Autor(a) principal: | |
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Data de Publicação: | 2002 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Repositório Institucional da UFSCAR |
Texto Completo: | https://repositorio.ufscar.br/handle/ufscar/5554 |
Resumo: | Phosphoribosylpyrophosphate synthetase (PRS - EC: 2.7.6.1) is an enzyme of central importance in several metabolic pathways in all cells. So far the plant PRS enzymes have not been investigated in great detail. However, accumulated evidences indicate that those enzymes form a complex family of isoenzymes with subcellular localization (mitochondria, cytoplasm and nucleus) and different phosphate dependence characteristics. The prs gene was cloned and had the sequence determined from a sugarcane cDNA library clone identified by the plant genome effort (SUCEST). The sugarcane prs gene contains a 984 bp open reading frame that encodes 328 amino acids protein with a calculated molecular weight of 36.6 kDa. The predicted amino acid sequence has 77 and 78% amino acid identity to the PRS4 of Arabidopsis thaliana and Spinacia oleracea respectively. The phylogenetic reconstruction of selected PRS homologues indicates that this enzyme may be a phosphate-independent PRS isoenzyme. The PRS protein was expressed in Escherichia coli, purified to homogeneity and found to retain secondary structure elements and quaternary arrangement consistent with the known PRS homologues. The availability of the PRS enzyme from another plant and the possibility of expressing the protein in large quantities should provide the basis for a functional and structural analysis of this important enzyme. |
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Sculaccio, Susana AndréaThiemann, Otávio Henriquehttp://genos.cnpq.br:12010/dwlattes/owa/prc_imp_cv_int?f_cod=K4788564Y19d9532eb-60ac-4962-8e57-85d7d3890e622016-06-02T20:21:37Z2004-08-132016-06-02T20:21:37Z2002-11-22https://repositorio.ufscar.br/handle/ufscar/5554Phosphoribosylpyrophosphate synthetase (PRS - EC: 2.7.6.1) is an enzyme of central importance in several metabolic pathways in all cells. So far the plant PRS enzymes have not been investigated in great detail. However, accumulated evidences indicate that those enzymes form a complex family of isoenzymes with subcellular localization (mitochondria, cytoplasm and nucleus) and different phosphate dependence characteristics. The prs gene was cloned and had the sequence determined from a sugarcane cDNA library clone identified by the plant genome effort (SUCEST). The sugarcane prs gene contains a 984 bp open reading frame that encodes 328 amino acids protein with a calculated molecular weight of 36.6 kDa. The predicted amino acid sequence has 77 and 78% amino acid identity to the PRS4 of Arabidopsis thaliana and Spinacia oleracea respectively. The phylogenetic reconstruction of selected PRS homologues indicates that this enzyme may be a phosphate-independent PRS isoenzyme. The PRS protein was expressed in Escherichia coli, purified to homogeneity and found to retain secondary structure elements and quaternary arrangement consistent with the known PRS homologues. The availability of the PRS enzyme from another plant and the possibility of expressing the protein in large quantities should provide the basis for a functional and structural analysis of this important enzyme.Fosforribosilpirofosfato sintetase (PRS EC: 2.7.6.1) é uma enzima de central importância em muitas vias metabólicas em todas as células. A PRS de plantas não tem sido investigada em grandes detalhes. Entretanto, as evidências acumuladas indicam que estas enzimas formam uma família complexa de isoenzimas com localização subcelular (mitocôndria, citoplasma e núcleo) e diferentes características de dependência a fosfato. O gene prs foi clonado e a seqüência determinada a partir de uma biblioteca de cDNA da cana-de-açúcar identificadas por um genoma de planta (SUCEST). O gene prs da cana-de-açúcar contem uma fase aberta de leitura de 984 pb que codifica uma proteína de 328 aminoácidos com massa molecular de 36,6 kDa. A seqüência de aminoácidos deduzida tem 77 e 78% de identidade coma PRS4 de Arabidopsis thaliana e Spinacia oleracea. A reconstrução filogenética de PRS homólogas selecionadas indica que esta enzima pode ser uma PRS indepentente de fosfato. Uma proteína foi expressa em Escherichia coli, purificada e encontrado os elementos de estrutura secundária e arranjos quaternários, consistentes com PRS homólogas conhecidas. A disponibilidade da enzima PRS a partir de uma outra planta e a possibilidade de expressar a proteína em larga escala pode fornecer a base para análises funcionais e estruturais desta importante enzima.application/pdfporUniversidade Federal de São CarlosPrograma de Pós-Graduação em Genética Evolutiva e Biologia Molecular - PPGGEvUFSCarBRBiologia molecularClonagemBiosíntesePurinasFosforribosil pirofosfatoCIENCIAS BIOLOGICAS::GENETICAClonagem e caracterização molecular da fosforribosil pirofosfato sintase (PRPP sintase) de cana-de-açucar.info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesis-1-1cc6c7897-1c5e-41af-8e84-9c960b60ad3binfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFSCARinstname:Universidade Federal de São Carlos (UFSCAR)instacron:UFSCARORIGINALDissSAS.pdfapplication/pdf3906877https://repositorio.ufscar.br/bitstream/ufscar/5554/1/DissSAS.pdf3cd2a799eac951c41dcb2ed83da21691MD51THUMBNAILDissSAS.pdf.jpgDissSAS.pdf.jpgIM Thumbnailimage/jpeg9166https://repositorio.ufscar.br/bitstream/ufscar/5554/2/DissSAS.pdf.jpg2eea65e76757141901355793e28051deMD52ufscar/55542023-09-18 18:31:07.324oai:repositorio.ufscar.br:ufscar/5554Repositório InstitucionalPUBhttps://repositorio.ufscar.br/oai/requestopendoar:43222023-09-18T18:31:07Repositório Institucional da UFSCAR - Universidade Federal de São Carlos (UFSCAR)false |
dc.title.por.fl_str_mv |
Clonagem e caracterização molecular da fosforribosil pirofosfato sintase (PRPP sintase) de cana-de-açucar. |
title |
Clonagem e caracterização molecular da fosforribosil pirofosfato sintase (PRPP sintase) de cana-de-açucar. |
spellingShingle |
Clonagem e caracterização molecular da fosforribosil pirofosfato sintase (PRPP sintase) de cana-de-açucar. Sculaccio, Susana Andréa Biologia molecular Clonagem Biosíntese Purinas Fosforribosil pirofosfato CIENCIAS BIOLOGICAS::GENETICA |
title_short |
Clonagem e caracterização molecular da fosforribosil pirofosfato sintase (PRPP sintase) de cana-de-açucar. |
title_full |
Clonagem e caracterização molecular da fosforribosil pirofosfato sintase (PRPP sintase) de cana-de-açucar. |
title_fullStr |
Clonagem e caracterização molecular da fosforribosil pirofosfato sintase (PRPP sintase) de cana-de-açucar. |
title_full_unstemmed |
Clonagem e caracterização molecular da fosforribosil pirofosfato sintase (PRPP sintase) de cana-de-açucar. |
title_sort |
Clonagem e caracterização molecular da fosforribosil pirofosfato sintase (PRPP sintase) de cana-de-açucar. |
author |
Sculaccio, Susana Andréa |
author_facet |
Sculaccio, Susana Andréa |
author_role |
author |
dc.contributor.author.fl_str_mv |
Sculaccio, Susana Andréa |
dc.contributor.advisor1.fl_str_mv |
Thiemann, Otávio Henrique |
dc.contributor.advisor1Lattes.fl_str_mv |
http://genos.cnpq.br:12010/dwlattes/owa/prc_imp_cv_int?f_cod=K4788564Y1 |
dc.contributor.authorID.fl_str_mv |
9d9532eb-60ac-4962-8e57-85d7d3890e62 |
contributor_str_mv |
Thiemann, Otávio Henrique |
dc.subject.por.fl_str_mv |
Biologia molecular Clonagem Biosíntese Purinas Fosforribosil pirofosfato |
topic |
Biologia molecular Clonagem Biosíntese Purinas Fosforribosil pirofosfato CIENCIAS BIOLOGICAS::GENETICA |
dc.subject.cnpq.fl_str_mv |
CIENCIAS BIOLOGICAS::GENETICA |
description |
Phosphoribosylpyrophosphate synthetase (PRS - EC: 2.7.6.1) is an enzyme of central importance in several metabolic pathways in all cells. So far the plant PRS enzymes have not been investigated in great detail. However, accumulated evidences indicate that those enzymes form a complex family of isoenzymes with subcellular localization (mitochondria, cytoplasm and nucleus) and different phosphate dependence characteristics. The prs gene was cloned and had the sequence determined from a sugarcane cDNA library clone identified by the plant genome effort (SUCEST). The sugarcane prs gene contains a 984 bp open reading frame that encodes 328 amino acids protein with a calculated molecular weight of 36.6 kDa. The predicted amino acid sequence has 77 and 78% amino acid identity to the PRS4 of Arabidopsis thaliana and Spinacia oleracea respectively. The phylogenetic reconstruction of selected PRS homologues indicates that this enzyme may be a phosphate-independent PRS isoenzyme. The PRS protein was expressed in Escherichia coli, purified to homogeneity and found to retain secondary structure elements and quaternary arrangement consistent with the known PRS homologues. The availability of the PRS enzyme from another plant and the possibility of expressing the protein in large quantities should provide the basis for a functional and structural analysis of this important enzyme. |
publishDate |
2002 |
dc.date.issued.fl_str_mv |
2002-11-22 |
dc.date.available.fl_str_mv |
2004-08-13 2016-06-02T20:21:37Z |
dc.date.accessioned.fl_str_mv |
2016-06-02T20:21:37Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/masterThesis |
format |
masterThesis |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
https://repositorio.ufscar.br/handle/ufscar/5554 |
url |
https://repositorio.ufscar.br/handle/ufscar/5554 |
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por |
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por |
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-1 -1 |
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cc6c7897-1c5e-41af-8e84-9c960b60ad3b |
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info:eu-repo/semantics/openAccess |
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openAccess |
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application/pdf |
dc.publisher.none.fl_str_mv |
Universidade Federal de São Carlos |
dc.publisher.program.fl_str_mv |
Programa de Pós-Graduação em Genética Evolutiva e Biologia Molecular - PPGGEv |
dc.publisher.initials.fl_str_mv |
UFSCar |
dc.publisher.country.fl_str_mv |
BR |
publisher.none.fl_str_mv |
Universidade Federal de São Carlos |
dc.source.none.fl_str_mv |
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