|
rdf:type |
|
|
lifeskim:mentions |
|
|
pubmed:issue |
2
|
|
pubmed:dateCreated |
2001-8-7
|
|
pubmed:databankReference |
|
|
pubmed:abstractText |
A cDNA clone (Atakn1) from Arabidopsis thaliana encoding APS-kinase (EC 2.7.1.25) was investigated for structural and catalytic properties of the gene product. Recombinant his10-AtAkn1 formed PAPS at a Vmax of 7.35 U x mg(-1). The Km for APS was 0.14 microM and for ATP 147 microM. APS caused a severe substrate inhibition (K(i) 4.5 microM). The type of inhibition is uncompetitive with respect to MgATP. High ionic strength and reducing thiols stabilized the enzyme activity. Plant APS-kinase is regulated in vitro by the redox charge with thioredoxin as essential activator. Mutagenesis of a serine in S182C and S182F presumed to be involved in the transfer of the phosphoryl group had no effect upon catalytic activity. Using a yeast two-hybrid system with AtAkn1 as bait, an interacting clone was detected from a cDNA library of A. thaliana cv. Columbia that codes for an APS-kinase iso-form (Atakn2). Complementation of APS-kinase-deficient Saccharomyces cerevisiae met14 showed that AtAkn2 is functionally active as APS-kinase. It was immunologically related to AtAkn1 and presumably represents a plastidal iso-form of the plant APS-kinase gene family.
|
|
pubmed:language |
eng
|
|
pubmed:journal |
|
|
pubmed:citationSubset |
IM
|
|
pubmed:chemical |
|
|
pubmed:status |
MEDLINE
|
|
pubmed:month |
Aug
|
|
pubmed:issn |
0003-9861
|
|
pubmed:author |
|
|
pubmed:copyrightInfo |
Copyright 2001 Academic Press.
|
|
pubmed:issnType |
Print
|
|
pubmed:day |
15
|
|
pubmed:volume |
392
|
|
pubmed:owner |
NLM
|
|
pubmed:authorsComplete |
Y
|
|
pubmed:pagination |
303-10
|
|
pubmed:dateRevised |
2007-11-15
|
|
pubmed:meshHeading |
pubmed-meshheading:11488606-Amino Acid Sequence,
pubmed-meshheading:11488606-Arabidopsis,
pubmed-meshheading:11488606-Binding, Competitive,
pubmed-meshheading:11488606-Catalysis,
pubmed-meshheading:11488606-Chromatography, High Pressure Liquid,
pubmed-meshheading:11488606-DNA, Complementary,
pubmed-meshheading:11488606-Dose-Response Relationship, Drug,
pubmed-meshheading:11488606-Electrophoresis, Polyacrylamide Gel,
pubmed-meshheading:11488606-Genetic Complementation Test,
pubmed-meshheading:11488606-Immunoblotting,
pubmed-meshheading:11488606-Kinetics,
pubmed-meshheading:11488606-Molecular Sequence Data,
pubmed-meshheading:11488606-Mutagenesis, Site-Directed,
pubmed-meshheading:11488606-Oxidation-Reduction,
pubmed-meshheading:11488606-Oxidoreductases,
pubmed-meshheading:11488606-Phosphotransferases (Alcohol Group Acceptor),
pubmed-meshheading:11488606-Protein Isoforms,
pubmed-meshheading:11488606-Recombinant Proteins,
pubmed-meshheading:11488606-Serine,
pubmed-meshheading:11488606-Thioredoxins,
pubmed-meshheading:11488606-Time Factors,
pubmed-meshheading:11488606-Two-Hybrid System Techniques
|
|
pubmed:year |
2001
|
|
pubmed:articleTitle |
Molecular and catalytic properties of Arabidopsis thaliana adenylyl sulfate (APS)-kinase.
|
|
pubmed:affiliation |
Biochemistry of Plants, Ruhr-University Bochum, Bochum, 44780, Germany.
|
|
pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
|
