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dc.contributor.author
Yep, Alejandra
dc.contributor.author
Bejar, Clarisa M.
dc.contributor.author
Ballicora, Miguel A.
dc.contributor.author
Dubay, Jennifer R.
dc.contributor.author
Iglesias, Alberto Alvaro
dc.contributor.author
Preiss, Jack
dc.date.available
2019-10-11T15:33:56Z
dc.date.issued
2004-01
dc.identifier.citation
Yep, Alejandra; Bejar, Clarisa M.; Ballicora, Miguel A.; Dubay, Jennifer R.; Iglesias, Alberto Alvaro; et al.; An assay for adenosine 5′-diphosphate (ADP)-glucose pyrophosphorylase that measures the synthesis of radioactive ADP-glucose with glycogen synthase; Academic Press Inc Elsevier Science; Analytical Biochemistry; 324; 1; 1-2004; 52-59
dc.identifier.issn
0003-2697
dc.identifier.uri
http://hdl.handle.net/11336/85713
dc.description.abstract
Adenosine 5′-diphosphate (ADP)-glucose pyrophosphorylase (ADP-Glc PPase) catalyzes the conversion of glucose 1-phosphate and adenosine 5′-triphosphate to ADP-glucose and pyrophosphate. We present a radioactive assay of this enzyme with a higher signal/noise ratio. After stopping the reaction that uses [14C]glucose 1-phosphate as a substrate, the ADP-[14C]glucose formed as a product is converted to [14C]glycogen by the addition of glycogen synthase and nonradioactive glycogen as primer. The final product is precipitated and washed, and the radioactivity is measured in a scintillation counter. The [ 14C]glucose 1-phosphate that did not react is easily eliminated during the washes. We have found that this assay produces much lower blanks than previously described radioactive methods based on binding of ADP-[ 14C]glucose to O-(diethylaminoethyl)-cellulose paper. In addition, we tested the kinetic parameters for the effectors of the Escherichia coli ADP-Glc PPase and both assays yielded identical results. The presented method is more suitable for Km or S0.5 determinations of ADP-Glc PPases having high apparent affinity for glucose 1-phosphate. It is possible to use a higher specific radioactivity to increase the sensitivity at lower concentrations of [14C]glucose 1-phosphate without compromising the blanks obtained at higher concentrations.
dc.format
application/pdf
dc.language.iso
eng
dc.publisher
Academic Press Inc Elsevier Science
dc.rights
info:eu-repo/semantics/restrictedAccess
dc.rights.uri
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.subject
ADP-GLUCOSE
dc.subject
ADP-GLUCOSE PYROPHOSPHORYLASE
dc.subject
GLYCOGEN SYNTHASE
dc.subject
POLYSACCHARIDE PRECIPITATION
dc.subject.classification
Bioquímica y Biología Molecular
dc.subject.classification
Ciencias Biológicas
dc.subject.classification
CIENCIAS NATURALES Y EXACTAS
dc.title
An assay for adenosine 5′-diphosphate (ADP)-glucose pyrophosphorylase that measures the synthesis of radioactive ADP-glucose with glycogen synthase
dc.type
info:eu-repo/semantics/article
dc.type
info:ar-repo/semantics/artículo
dc.type
info:eu-repo/semantics/publishedVersion
dc.date.updated
2019-10-10T19:34:19Z
dc.journal.volume
324
dc.journal.number
1
dc.journal.pagination
52-59
dc.journal.pais
Estados Unidos
dc.description.fil
Fil: Yep, Alejandra. Michigan State University; Estados Unidos
dc.description.fil
Fil: Bejar, Clarisa M.. Michigan State University; Estados Unidos
dc.description.fil
Fil: Ballicora, Miguel A.. Michigan State University; Estados Unidos
dc.description.fil
Fil: Dubay, Jennifer R.. Michigan State University; Estados Unidos
dc.description.fil
Fil: Iglesias, Alberto Alvaro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Agrobiotecnología del Litoral. Universidad Nacional del Litoral. Instituto de Agrobiotecnología del Litoral; Argentina
dc.description.fil
Fil: Preiss, Jack. Michigan State University; Estados Unidos
dc.journal.title
Analytical Biochemistry
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/doi/http://dx.doi.org/10.1016/j.ab.2003.09.024


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