Geranyl diphosphate synthase: Cloning, expression, and characterization of this prenyltransferase as a heterodimer

Abstract

Geranyl diphosphate synthase, which catalyzes the condensation of dimethylallyl diphosphate and isopentenyl diphosphate to geranyl diphosphate, the key precursor of monoterpene biosynthesis, was purified from isolated oil glands of spearmint. Peptide fragments generated from the pure proteins of 28 and 37 kDa revealed amino acid sequences that matched two cDNA clones obtained by random screening of a peppermint-oil gland cDNA library. The deduced sequences of both proteins showed some similarity to existing prenyltransferases, and both contained a plastid-targeting sequence. Expression of each cDNA individually yielded no detectable prenyltransferase activity; however, coexpression of the two together produced functional geranyl diphosphate synthase. Antibodies raised against each protein were used to demonstrate that both subunits were required to produce catalytically active native and recombinant enzymes, thus confirming that geranyl diphosphate synthase is a heterodimer. monoterpene biosynthesis ͉ isoprenoid biosynthesis

Five Geranylgeranyl Diphosphate Synthases Expressed in Different Organs Are Localized into Three Subcellular Compartments in Arabidopsis

“…8). Furthermore, the large subunit of GPP synthase from peppermint has considerable similarity to Arabidopsis GGR protein (Burke et al, 1999), suggesting that the GGR gene would encode the GPP synthase large subunit rather than GGPP synthase.…”

Induction of a leaf specific geranylgeranyl pyrophosphate synthase and emission of (E,E)-4,8,12-trimethyltrideca-1,3,7,11-tetraene in tomato are dependent on both jasmonic acid and salicylic acid signaling pathways

“…This system has been used before to demonstrate the function of putative GGPP synthases from Arabidopsis (Zhu et al 1997), sunXower (Helianthus annuus) (Oh et al 2000) makandi (Coleus forskohlii) (Engprasert et al 2004), human and mouse (Mus musculus) (Kainou et al 1999). While GGPP synthase, FPP synthase and most GPP synthases are functional as homodimers (Ogura and Koyama 1998), the GPP synthases from mint and snapdragon are functional as heterodimers (Burke et al 1999;Tholl et al 2004). In the phylogenetic tree ( Fig.…”

Heterodimeric geranyl(geranyl)diphosphate synthase from hop ( Humulus lupulus ) and the evolution of monoterpene biosynthesis

“…Heterodimeric GPPSs have been described from angiosperms (5,6), and homodimeric GPPSs have been functionally characterized from both angiosperms (7,8) and gymnosperms (9,10). Both subunits of homodimeric GPPSs and the large subunit of heterodimeric GPPSs (GPPS.LSU) contain 2 aspartate-rich motifs, DD(X) 2-4 D (where ''X'' is any amino acid), which are important in prenyl-substrate binding (11).…”