Human Geranylgeranyl Diphosphate Synthase is an Octamer in Solution

Miyagi, Yukino; Matsumura, Yoshihiro; Sagami, Hiroshi

doi:10.1093/jb/mvm144

Cited by 19 publications

(13 citation statements)

References 12 publications

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“…S4). For comparison, the human FPPS, the cotton boll weevil FPPS, and the horseradish leaf beetle GPPS/FPPS are dimers (11,12,16); human GGPPS is an octamer (17); and bovine GGPPS is a multimer of four to five units (18).…”

Section: Resultsmentioning

confidence: 99%

Frontalin pheromone biosynthesis in the mountain pine beetle, Dendroctonus ponderosae , and the role of isoprenyl diphosphate synthases

Keeling

Chiu

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

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The mountain pine beetle (Dendroctonus ponderosae Hopkins) is the most destructive pest of western North American pine forests. Adult males produce frontalin, an eight-carbon antiaggregation pheromone, via the mevalonate pathway, as part of several pheromones that initiate and modulate the mass attack of host trees. Frontalin acts as a pheromone, attractant, or kairomone in most Dendroctonus species, other insects, and even elephants. 6-Methylhept-6-en-2-one, a frontalin precursor, is hypothesized to originate from 10-carbon geranyl diphosphate (GPP), 15-carbon farnesyl diphosphate (FPP), or 20-carbon geranylgeranyl diphosphate (GGPP) via a dioxygenase-or cytochrome P450-mediated carboncarbon bond cleavage. To investigate the role of isoprenyl diphosphate synthases in pheromone biosynthesis, we characterized a bifunctional GPP/FPP synthase and a GGPP synthase in the mountain pine beetle. The ratio of GPP to FPP produced by the GPP/FPP synthase was highly dependent on the ratio of the substrates isopentenyl diphosphate and dimethylallyl diphosphate used in the assay. Transcript levels in various tissues and life stages suggested that GGPP rather than GPP or FPP is used as a precursor to frontalin. Reduction of transcript levels by RNA interference of the isoprenyl diphosphate synthases identified GGPP synthase as having the largest effect on frontalin production, suggesting that frontalin is derived from a 20-carbon isoprenoid precursor rather than from the 10-or 15-carbon precursors. (Fig. 1), is an antiaggregation pheromone known to be synthesized de novo via the mevalonate pathway in male midgut tissue (3, 4). Frontalin acts as a pheromone, attractant, or kairomone in most other Dendroctonus species, other insects (5), and even elephants (6, 7) and has been studied in bark beetles for more than 40 y (8). Its biosynthetic precursor, 6-methylhept-6-en-2-one (6MHO), found in both insects and elephants (6, 9), is hypothesized to originate from cleavage of geranyl diphosphate (GPP), farnesyl diphosphate (FPP), or geranylgeranyl diphosphate (GGPP) by a dioxygenase or cytochrome P450 (10). These isoprenyl diphosphates are biosynthesized from the five-carbon precursors isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP) by the isoprenyl diphosphate synthases (IDSs) GPP synthase (GPPS), FPP synthase (FPPS), and GGPP synthase (GGPPS). However, it is not known which of these isoprenyl diphosphates is used as a precursor in frontalin biosynthesis.A few IDSs have been characterized in Coleoptera, including an FPPS from the cotton boll weevil (Anthonomus grandis) (11), a GPPS/FPPS from the horseradish leaf beetle (Phaedon cochleariae) (12), and a unique GPPS from the pine engraver beetle (Ips pini) that also produces myrcene (13,14). To assess their importance in pheromone biosynthesis, we identified and functionally characterized two IDSs in MPB. Based on three different lines of evidence, gene-expression patterns of the IDSs, enzyme activity of recombinant proteins, and the frontalin content of dsRNA-trea...

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Section: Resultsmentioning

confidence: 99%

Frontalin pheromone biosynthesis in the mountain pine beetle, Dendroctonus ponderosae , and the role of isoprenyl diphosphate synthases

Keeling

Chiu

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

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“…Work done by Kavanagh et al (2006) suggested that based on molecular weight, 5-6 monomers are associated per molecule (molecular weight 193 kDa) in solution. Sagami et al (1994) reported that GGDPS from bovine brain is a homooligomer with a monomer molecular mass of 37.5 kDa, whereas Miyagi et al (2007) reported that the active form of GGDPS in solution is an octomer and that hexamer and dimer forms On the basis of the computational modeling studies, both HG and HN can simultaneously bind to distinct but overlapping sites within the enzyme (HG with preference for the GGDP site and HN with preference for the FDP site) (Fig. 8).…”

Section: Discussionmentioning

confidence: 99%

Olefin Isomers of a Triazole Bisphosphonate Synergistically Inhibit Geranylgeranyl Diphosphate Synthase

Allen

Kortagere

Tong

et al. 2017

Molecular Pharmacology

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The isoprenoid donor for protein geranylgeranylation reactions, geranylgeranyl diphosphate (GGDP), is the product of the enzyme GGDP synthase (GGDPS) that condenses farnesyl diphosphate (FDP) and isopentenyl pyrophosphate. GGDPS inhibition is of interest from a therapeutic perspective for multiple myeloma because we have shown that targeting Rab GTPase geranylgeranylation impairs monoclonal protein trafficking, leading to endoplasmic reticulum stress and apoptosis. We reported a series of triazole bisphosphonate GGDPS inhibitors, of which the most potent was a 3:1 mixture of homogeranyl (HG) and homoneryl (HN) isomers. Here we determined the activity of the individual olefin isomers. Enzymatic and cellular assays revealed that although HN is approximately threefold more potent than HG, HN is not more potent than the original mixture. Studies in which cells were treated with varying concentrations of each isomer alone and in different combinations revealed that the two isomers potentiate the induced-inhibition of protein geranylgeranylation when used in a 3:1 HG:HN combination. A synergistic interaction was observed between the two isomers in the GGDPS enzyme assay. These results suggested that the two isomers bind simultaneously to the enzyme but within different domains. Computational modeling studies revealed that HN is preferred at the FDP site, that HG is preferred at the GGDP site, and that both isomers may bind to the enzyme simultaneously. These studies are the first to report a set of olefin isomers that synergistically inhibit GGDPS, thus establishing a new paradigm for the future development of GGDPS inhibitors.

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“…However, three dimers of this enzyme join together to form a homohexamer complex, which has been described as a three-blade propeller (Figure 3B). Biochemical investigations have even described the formation of octameric complexes in solution (Miyagi et al, 2007). Despite low sequence identity (17%), the tertiary structure of hGGPPS is remarkably similar to that of hFPPS (Figure 3A).…”

Section: Human Ggpps—overall Structurementioning

confidence: 99%

Human isoprenoid synthase enzymes as therapeutic targets

et al. 2014

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In the human body, the complex biochemical network known as the mevalonate pathway is responsible for the biosynthesis of all isoprenoids, which consists of a vast array of metabolites that are vital for proper cellular functions. Two key isoprenoids, farnesyl pyrophosphate (FPP) and geranylgeranyl pyrophosphate (GGPP) are responsible for the post-translational prenylation of small GTP-binding proteins, and serve as the biosynthetic precursors to numerous other biomolecules. The down-stream metabolite of FPP and GGPP is squalene, the precursor to steroids, bile acids, lipoproteins, and vitamin D. In the past, interest in prenyl synthase inhibitors focused mainly on the role of the FPP in lytic bone diseases. More recently pre-clinical and clinical studies have strongly implicated high levels of protein prenylation in a plethora of human diseases, including non-skeletal cancers, the progression of neurodegenerative diseases and cardiovascular diseases. In this review, we focus mainly on the potential therapeutic value of down-regulating the biosynthesis of FPP, GGPP, and squalene. We summarize the most recent drug discovery efforts and the structural data available that support the current on-going studies.

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Human Geranylgeranyl Diphosphate Synthase is an Octamer in Solution

Cited by 19 publications

References 12 publications

Frontalin pheromone biosynthesis in the mountain pine beetle, Dendroctonus ponderosae , and the role of isoprenyl diphosphate synthases

Frontalin pheromone biosynthesis in the mountain pine beetle, Dendroctonus ponderosae , and the role of isoprenyl diphosphate synthases

Olefin Isomers of a Triazole Bisphosphonate Synergistically Inhibit Geranylgeranyl Diphosphate Synthase

Human isoprenoid synthase enzymes as therapeutic targets

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