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dc.contributor.author Asención Diez, Matías Damián
dc.contributor.author Ebrecht, Ana Cristina
dc.contributor.author Martínez, Lucila Inés
dc.contributor.author Aleanzi, Mabel Cristina
dc.contributor.author Guerrero, Sergio Adrian
dc.contributor.author Ballicora, Miguel A.
dc.contributor.author Iglesias, Alberto Alvaro
dc.date.available 2015-08-11T20:43:08Z
dc.date.issued 2013-05
dc.identifier.citation Asención Diez, Matías Damián; Ebrecht, Ana Cristina; Martínez, Lucila Inés; Aleanzi, Mabel Cristina; Guerrero, Sergio Adrian; et al.; A Chimeric UDP-Glucose Pyrophosphorylase Produced by Protein Engineering Exhibits Sensitivity to Allosteric Regulators; Molecular Diversity Preservation International; International Journal Of Molecular Sciences; 14; 5; 5-2013; 9703-9721
dc.identifier.issn 1422-0067
dc.identifier.uri http://hdl.handle.net/11336/1621
dc.description.abstract In bacteria, glycogen or oligosaccharide accumulation involves glucose-1-phosphate partitioning into either ADP-glucose (ADP-Glc) or UDP-Glc. Their respective synthesis is catalyzed by allosterically regulated ADP-Glc pyrophosphorylase (EC 2.7.7.27, ADP-Glc PPase) or unregulated UDP-Glc PPase (EC 2.7.7.9). In this work, we characterized the UDP-Glc PPase from Streptococcus mutans . In addition, we constructed a chimeric protein by cutting the C-terminal domain of the ADP-Glc PPase from Escherichia coli and pasting it to the entire S. mutans UDP-Glc PPase. Both proteins were fully active as UDP-Glc PPases and their kinetic parameters were measured. The chimeric enzyme had a slightly higher affinity for substrates than the native S. mutans UDP-Glc PPase, but the maximal activity was four times lower. Interestingly, the chimeric protein was sensitive to regulation by pyruvate, 3-phosphoglyceric acid and fructose-1,6-bis-phosphate, which are known to be effectors of ADP-Glc PPases from different sources. The three compounds activated the chimeric enzyme up to three-fold, and increased the affinity for substrates. This chimeric protein is the first reported UDP-Glc PPase with allosteric regulatory properties. In addition, this is a pioneer work dealing with a chimeric enzyme constructed as a hybrid of two pyrophosphorylases with different specificity toward nucleoside-diphospho-glucose and our results turn to be relevant for a deeper understanding of the evolution of allosterism in this family of enzymes.
dc.format application/pdf
dc.language.iso eng
dc.publisher Molecular Diversity Preservation International
dc.rights info:eu-repo/semantics/openAccess
dc.rights.uri https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.subject protein engineering
dc.subject allosteric regulation
dc.subject pyrophosphorylases evolution
dc.subject UDP-glucose
dc.subject ADP-glucose
dc.subject.classification Bioquímica y Biología Molecular
dc.subject.classification Ciencias Biológicas
dc.subject.classification CIENCIAS NATURALES Y EXACTAS
dc.title A Chimeric UDP-Glucose Pyrophosphorylase Produced by Protein Engineering Exhibits Sensitivity to Allosteric Regulators
dc.type info:eu-repo/semantics/article
dc.type info:ar-repo/semantics/artículo
dc.type info:eu-repo/semantics/publishedVersion
dc.date.updated 2016-03-30 10:35:44.97925-03
dc.journal.volume 14
dc.journal.number 5
dc.journal.pagination 9703-9721
dc.journal.pais Suiza
dc.journal.ciudad Basel
dc.description.fil Fil: Asención Diez, Matías Damián. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico - CONICET - Instituto de Agrobiotecnologia del Litoral; Argentina;
dc.description.fil Fil: Ebrecht, Ana Cristina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico - CONICET - Santa Fe. Instituto de Agrobiotecnologia del Litoral; Argentina;
dc.description.fil Fil: Martínez, Lucila Inés. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico - CONICET - Santa Fe. Instituto de Agrobiotecnologia del Litoral; Argentina;
dc.description.fil Fil: Aleanzi, Mabel Cristina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico - CONICET - Santa Fe. Instituto de Agrobiotecnologia del Litoral; Argentina;
dc.description.fil Fil: Guerrero, Sergio Adrian. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico - CONICET - Santa Fe. Instituto de Agrobiotecnologia del Litoral; Argentina;
dc.description.fil Fil: Ballicora, Miguel A.. University Of Chicago; Estados Unidos de América;
dc.description.fil Fil: Iglesias, Alberto Alvaro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico - CONICET - Santa Fe. Instituto de Agrobiotecnologia del Litoral; Argentina;
dc.journal.title International Journal Of Molecular Sciences
dc.relation.alternativeid info:eu-repo/semantics/altIdentifier/doi/http://dx.doi.org/10.3390/ijms14059703
dc.relation.alternativeid info:eu-repo/semantics/altIdentifier/url/http://www.mdpi.com/1422-0067/14/5/9703


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info:eu-repo/semantics/openAccess Excepto donde se diga explícitamente, este item se publica bajo la siguiente descripción: Creative Commons Attribution-NonCommercial-ShareAlike 2.5 Unported (CC BY-NC-SA 2.5)