Linked Life Data

19968859

Source:http://linkedlifedata.com/resource/pubmed/id/19968859

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
2
pubmed:dateCreated
2010-1-12
pubmed:abstractText
Starch synthase III from Arabidopsis thaliana contains an N-terminal region, including three in-tandem starch-binding domains, followed by a C-terminal catalytic domain. We have reported previously that starch-binding domains may be involved in the regulation of starch synthase III function. In this work, we analyzed the existence of protein interactions between both domains using pull-down assays, far western blotting and co-expression of the full and truncated starch-binding domains with the catalytic domain. Pull-down assays and co-purification analysis showed that the D(316-344) and D(495-535) regions in the D2 and D3 domains, respectively, but not the individual starch-binding domains, are involved in the interaction with the catalytic domain. We also determined that the residues W366 and Y394 in the D2 domain are important in starch binding. Moreover, the co-purified catalytic domain plus site-directed mutants of the D123 protein lacking these aromatic residues showed that W366 was key to the apparent affinity for the polysaccharide substrate of starch synthase III, whereas either of these amino acid residues altered ADP-glucose kinetics. In addition, the analysis of full-length and truncated proteins showed an almost complete restoration of the apparent affinity for the substrates and V(max) of starch synthase III. The results presented here suggest that the interaction of the N-terminal starch-binding domains, particularly the D(316-344) and D(495-535) regions, with the catalytic domains, as well as the full integrity of the starch-binding capacity of the D2 domain, are involved in the modulation of starch synthase III activity.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Jan
pubmed:issn
1742-4658
pubmed:author
pubmed:issnType
Electronic
pubmed:volume
277
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
428-40
pubmed:meshHeading
pubmed-meshheading:19968859-Amino Acid Sequence, pubmed-meshheading:19968859-Amino Acid Substitution, pubmed-meshheading:19968859-Arabidopsis, pubmed-meshheading:19968859-Arabidopsis Proteins, pubmed-meshheading:19968859-Base Sequence, pubmed-meshheading:19968859-Binding Sites, pubmed-meshheading:19968859-DNA, Plant, pubmed-meshheading:19968859-DNA Primers, pubmed-meshheading:19968859-Glucosyltransferases, pubmed-meshheading:19968859-Kinetics, pubmed-meshheading:19968859-Molecular Sequence Data, pubmed-meshheading:19968859-Mutagenesis, Site-Directed, pubmed-meshheading:19968859-Protein Interaction Domains and Motifs, pubmed-meshheading:19968859-Protein Structure, Tertiary, pubmed-meshheading:19968859-Recombinant Proteins, pubmed-meshheading:19968859-Starch, pubmed-meshheading:19968859-Substrate Specificity
pubmed:year
2010
pubmed:articleTitle
The starch-binding capacity of the noncatalytic SBD2 region and the interaction between the N- and C-terminal domains are involved in the modulation of the activity of starch synthase III from Arabidopsis thaliana.
pubmed:affiliation
Instituto de Investigaciones Biotecnológicas-Instituto Tecnológico de Chascomús, Argentina.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't