Molecular cloning and characterization of a prenyltransferase from the cotton aphid, Aphis gossypii

Ma, Guang-Yuan; Sun, Xiaohong; Zhang, Yong-Lei; Li, Zhengxi; Shen, Zuorui

doi:10.1016/j.ibmb.2010.05.003

Cited by 23 publications

(30 citation statements)

References 40 publications

(59 reference statements)

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“…Bifunctionality was also observed from the scIDSs characterized from different aphid species (13)(14)(15)(16)(17). Here, the recombinant proteins generated both GDP and FDP in parallel, and hence may be involved in the biosynthesis of either aphid sex pheromones or the sesquiterpene (E)-β-farnesene, the most common component of alarm pheromones.…”

mentioning

confidence: 70%

“…Our kinetic data for PcIDS1 support the observation that the regulation of product distribution by these metal cofactors is biochemically relevant. Comparisons , respectively, are in the range of K m values for previously described insect scIDSs, such as those of A. gossypii (12.6 μM) (14), D. melanogaster FPPS-1b (7 μM) (32), M. sexta (0.8 μM) (27), and M. persicae (MpIPPS1-S, 25.4 μM; MpIPPS2-S, 15.4 μM) (17). The difference in the PcIDS1 K m values for different metal cofactors suggests that the enzyme favors GDP as a substrate with Mg…”

Section: +mentioning

confidence: 99%

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Metal ions control product specificity of isoprenyl diphosphate synthases in the insect terpenoid pathway

Frick¹,

Nagel

Schmidt

et al. 2013

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

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Isoprenyl diphosphate synthases (IDSs) produce the ubiquitous branched-chain diphosphates of different lengths that are precursors of all major classes of terpenes. Typically, individual shortchain IDSs (scIDSs) make the C 10 , C 15 , and C 20 isoprenyl diphosphates separately. Here, we report that the product length synthesized by a single scIDS shifts depending on the divalent metal cofactor present. This previously undescribed mechanism of carbon chainlength determination was discovered for a scIDS from juvenile horseradish leaf beetles, Phaedon cochleariae. The recombinant enzyme P. cochleariae isoprenyl diphosphate synthase 1 (PcIDS1) yields 96% C 10 -geranyl diphosphate (GDP) and only 4% C 15 -farnesyl diphosphate (FDP) in the presence of Co 2+ or Mn 2+ as a cofactor, whereas it yields only 18% C 10 GDP but 82% C 15 FDP in the presence of Mg 2+. In reaction with Co 2+ , PcIDS1 has a K m of 11.6 μM for dimethylallyl diphosphate as a cosubstrate and 24.3 μM for GDP. However, with Mg 2+ , PcIDS1 has a K m of 1.18 μM for GDP, suggesting that this substrate is favored by the enzyme under such conditions. RNAi targeting PcIDS1 revealed the participation of this enzyme in the de novo synthesis of defensive monoterpenoids in the beetle larvae. As an FDP synthase, PcIDS1 could be associated with the formation of sesquiterpenes, such as juvenile hormones. Detection of Co , or Mg 2+ in the beetle larvae suggests flux control into C 10 vs. C 15 isoprenoids could be accomplished by these ions in vivo. The dependence of product chain length of scIDSs on metal cofactor identity introduces an additional regulation for these branch point enzymes of terpene metabolism.cobalt | kinetic | prenyltransferase | secretions | silencing

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mentioning

confidence: 70%

Section: +mentioning

confidence: 99%

Metal ions control product specificity of isoprenyl diphosphate synthases in the insect terpenoid pathway

Frick¹,

Nagel

Schmidt

et al. 2013

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

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“…bark beetles produced C 10 -GDP and C 15 -farnesyl diphosphate (C 15 -FDP) depending on the C 5 -IDP/C 5 -DMADP substrate ratio [79][80][81]. Bifunctionality was also observed from the scIDSs characterized from different aphid species [83][84][85][86][87]. Here, the recombinant proteins generated both GDP and FDP in parallel, and hence may be involved in the biosynthesis of either aphid sex pheromones or the sesquiterpene (E)-β-farnesene, the most common component of alarm pheromones.…”

Section: Iridoid De Novo Synthesis: Early Stepsmentioning

confidence: 99%

Deciphering the route to cyclic monoterpenes in Chrysomelina leaf beetles: source of new biocatalysts for industrial application?

Burse

Boland

2017

Zeitschrift Für Naturforschung C

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Abstract:The drastic growth of the population on our planet requires the efficient and sustainable use of our natural resources. Enzymes are indispensable tools for a wide range of industries producing food, pharmaceuticals, pesticides, or biofuels. Because insects constitute one of the most species-rich classes of organisms colonizing almost every ecological niche on earth, they have developed extraordinary metabolic abilities to survive in various and sometimes extreme habitats. Despite this metabolic diversity, insect enzymes have only recently generated interest in industrial applications because only a few metabolic pathways have been sufficiently characterized. Here, we address the biosynthetic route to iridoids (cyclic monoterpenes), a group of secondary metabolites used by some members of the leaf beetle subtribe Chrysomelina as defensive compounds against their enemies. The ability to produce iridoids de novo has also convergently evolved in plants. From plant sources, numerous pharmacologically relevant structures have already been described. In addition, in plants, iridoids serve as building blocks for monoterpenoid indole alkaloids with broad therapeutic applications. As the commercial synthesis of iridoidbased drugs often relies on a semisynthetic approach involving biocatalysts, the discovery of enzymes from the insect iridoid route can account for a valuable resource and economic alternative to the previously used enzymes from the metabolism of plants. Hence, this review illustrates the recent discoveries made on the steps of the iridoid pathway in Chrysomelina leaf beetles. The findings are also placed in the context of the studied counterparts in plants and are further discussed regarding their use in technological approaches.

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“…The procedure for cloning the cDNAs of the two aphid FPPSs (designated as RpFPPS1 and RpFPPS2) was similar to that previously described [13,28]. Briefly, first-strand cDNA was synthesized from total RNA at 42°C using oligo(dT) 18 primer and PowerScript reverse transcriptase (Clontech, USA), according to the manufacturer's protocol.…”

Section: The Aphids and Cdna Cloningmentioning

confidence: 99%

“…Other insect FPPSs have been identified by sequence homology in the past decade or so [4,8,12,13,20,25]. These studies have mainly been carried out with emphasis on gene cloning, sequence analysis and activity analysis.…”

Section: Introductionmentioning

confidence: 99%

In Silico and In Vitro Analyses Identified Three Amino Acid Residues Critical to the Catalysis of Two Aphid Farnesyl Diphosphate Synthase

Sun

2012

Protein J

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Farnesyl diphosphate synthase (FPPS) plays an essential role in the isoprenoid biosynthetic pathway of microbes, plants and animals. In the present study, we first cloned two FPPSs from the bird cherry-oat aphid (RpFPPS1 and RpFPPS2), and activity assay by gas chromatography-mass spectrometry showed that both RpFPPS1 and RpFPPS2 were active in vitro. They were then subjected to homology modeling and molecular docking. Molecular interaction analysis indicated that three amino acid residues (R120, R121 and K266) might play key roles in the catalysis of the two aphid FPPSs by forming hydrogen bonds with the diphosphate moiety of the allylic substrate. These in silico results were subsequently confirmed by site-directed mutagenesis and in vitro activity assay of the mutant enzymes, in which each of the single mutations R120G, R121G and K266I abolished the activities of the two FPPSs. This study contributes to our understanding of the catalytic mechanism of farnesyl diphosphate synthases.

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Molecular cloning and characterization of a prenyltransferase from the cotton aphid, Aphis gossypii

Cited by 23 publications

References 40 publications

Metal ions control product specificity of isoprenyl diphosphate synthases in the insect terpenoid pathway

Metal ions control product specificity of isoprenyl diphosphate synthases in the insect terpenoid pathway

Deciphering the route to cyclic monoterpenes in Chrysomelina leaf beetles: source of new biocatalysts for industrial application?

In Silico and In Vitro Analyses Identified Three Amino Acid Residues Critical to the Catalysis of Two Aphid Farnesyl Diphosphate Synthase

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