Cloning and expression of maize-leaf pyruvate, Pi dikinase regulatory protein gene.
Pyruvate, orthophosphate dikinase (PPDK; E.C. 2.7.9.1) catalyzes the synthesis of the primary inorganic carbon acceptor, phosphoenolpyruvate in the C4 photosynthetic pathway and is reversibly regulated by light. PPDK regulatory protein (RP), a bifunctional serine/threonine kinase-phosphatase, catalyzes both the ADP-dependent inactivation and the Pi-dependent activation of PPDK. Attempts to clone the RP have to date proven unsuccessful. A bioinformatics approach was taken to identify the nucleotide and amino acid sequence of the protein. Based on previously established characteristics including molecular mass, known inter- and intracellular location, functionality, and low level of expression, available databases were interrogated to ultimately identify a single candidate gene. In this paper, we describe the nucleotide and deduced amino acid sequence of this gene and establish its identity as maize PPDK RP by in vitro analysis of its catalytic properties via the cloning and expression of the recombinant protein.
Burnell
Jim N
JN
Department of Biochemistry and Molecular Biology, James Cook University, Townsville, Qld 4811, Australia. James.Burnell@jcu.edu.au
Chastain
Chris J
CJ
D000595:Amino Acid Sequence; D001483:Base Sequence; D002384:Catalysis; D019976:Cloning, Organism; D019295:Computational Biology; D005110:Extracellular Space / Q000378:metabolism; D018506:Gene Expression Regulation, Plant; D042541:Intracellular Space / Q000378:metabolism; D008969:Molecular Sequence Data; D018515:Plant Leaves / Q000235:genetics* / Q000378:metabolism; D011772:Pyruvate, Orthophosphate Dikinase / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D003313:Zea mays / Q000235:genetics*
D011994:Recombinant Proteins; D011772:Pyruvate, Orthophosphate Dikinase