Cloning and expression of maize-leaf pyruvate, Pi dikinase regulatory protein gene

• Published in: Biochemical and biophysical research communications 2006-06, Vol.345 (2), p.675-680
• Main Authors: Burnell, Jim N., Chastain, Chris J.
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Base Sequence; C 4 photosynthesis; Catalysis; Cloning, Organism; Computational Biology; Extracellular Space - metabolism; Gene Expression Regulation, Plant; Intracellular Space - metabolism; Maize; Molecular Sequence Data; Plant Leaves - genetics; Plant Leaves - metabolism; PPDK; PPDK regulatory protein; Protein phosphorylation/dephosphorylation; Pyruvate, Orthophosphate Dikinase - genetics; Pyruvate, Orthophosphate Dikinase - metabolism; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; Zea mays; Zea mays - genetics
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1016/j.bbrc.2006.04.150

Pyruvate, orthophosphate dikinase (PPDK; E.C. 2.7.9.1) catalyzes the synthesis of the primary inorganic carbon acceptor, phosphoenolpyruvate in the C 4 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.