Trinuclear metal clusters in catalysis by terpenoid synthases

Aaron, Julie A.; Christianson, D.W.

doi:10.1351/pac-con-09-09-37

Cited by 127 publications

(164 citation statements)

References 57 publications

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“…Typically, diphosphate or bisphosphonate binding to terpenoid synthases is accommodated by 3 metal ions and 3 basic residues, and these interactions ensure complete active site closure. 28 …”

Section: Resultsmentioning

confidence: 99%

Structure and Function of Fusicoccadiene Synthase, a Hexameric Bifunctional Diterpene Synthase

et al. 2016

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Fusicoccin A is a diterpene glucoside phytotoxin generated by the fungal pathogen Phomopsis amygdali that causes the plant disease constriction canker, first discovered in New Jersey peach orchards in the 1930’s. Fusicoccin A is also an emerging new lead in cancer chemotherapy. The hydrocarbon precursor of fusicoccin A is the tricyclic diterpene fusicoccadiene, which is generated by a bifunctional terpenoid synthase. Here, we report X-ray crystal structures of the individual catalytic domains of fusicoccadiene synthase: the C-terminal domain is a chain elongation enzyme that generates geranylgeranyl diphosphate, and the N-terminal domain catalyzes the cyclization of geranylgeranyl diphosphate to form fusicoccadiene. Crystal structures of each domain complexed with bisphosphonate substrate analogues suggest that three metal ions and three positively charged amino acid side chains trigger substrate ionization in each active site. While in vitro incubations reveal that the cyclase domain can utilize farnesyl diphosphate and geranyl diphosphate as surrogate substrates, these shorter isoprenoid diphosphates are mainly converted into acyclic alcohol or hydrocarbon products. Gel filtration chromatography and analytical ultracentrifugation experiments indicate that full-length fusicoccadiene synthase adopts hexameric quaternary structure, and small-angle X-ray scattering data yield a well-defined molecular envelope illustrating a plausible model for hexamer assembly.

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Section: Resultsmentioning

confidence: 99%

Structure and Function of Fusicoccadiene Synthase, a Hexameric Bifunctional Diterpene Synthase

et al. 2016

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“…However, a second conserved metal binding motif, NDXXSXXXE, which is characteristic of nearly all class I terpene synthases (Aaron and Christianson, 2010), is completely conserved in five out of eight sequences, MpMTPSL1-4 and 6. The corresponding sequence is apparent in the other MpMTPSL proteins, but missing or having substitutions for highly conserved residues (MpMTPSL5 and 8, G for S substitution; MpMTPSL7, S for N substitution).…”

Section: Identification Of Putative M Polymorpha Terpene Synthasesmentioning

confidence: 99%

Molecular Diversity of Terpene Synthases in the Liverwort Marchantia polymorpha

et al. 2016

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Marchantia polymorpha is a basal terrestrial land plant, which like most liverworts accumulates structurally diverse terpenes believed to serve in deterring disease and herbivory. Previous studies have suggested that the mevalonate and methylerythritol phosphate pathways, present in evolutionarily diverged plants, are also operative in liverworts. However, the genes and enzymes responsible for the chemical diversity of terpenes have yet to be described. In this study, we resorted to a HMMER search tool to identify 17 putative terpene synthase genes from M. polymorpha transcriptomes. Functional characterization identified four diterpene synthase genes phylogenetically related to those found in diverged plants and nine rather unusual monoterpene and sesquiterpene synthase-like genes. The presence of separate monofunctional diterpene synthases for ent-copalyl diphosphate and ent-kaurene biosynthesis is similar to orthologs found in vascular plants, pushing the date of the underlying gene duplication and neofunctionalization of the ancestral diterpene synthase gene family to >400 million years ago. By contrast, the mono-and sesquiterpene synthases represent a distinct class of enzymes, not related to previously described plant terpene synthases and only distantly so to microbial-type terpene synthases. The absence of a Mg 2+ binding, aspartate-rich, DDXXD motif places these enzymes in a noncanonical family of terpene synthases.

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“…The more than 55,000 terpenes identified thus far are crucial components of intracellular signal-transduction pathways, electron transport chains, and membranes, or they can function as hormones, photosynthetic pigments, and semiochemicals (1,2). Despite their structural diversity, terpenes are derived from the universal linear C 10 , C 15 , C 20 , and larger diphosphate intermediates whose synthesis is catalyzed by isoprenyl diphosphate synthases (IDSs), also known as prenyltransferases (3). Depending on the stereochemistry of the double bond of the reaction product, these enzymes are classified as either trans-IDSs or cisIDSs (4,5).…”

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confidence: 99%

“…During chain elongation, the allylic cosubstrate (DMADP or GDP) undergoes coupling with IDP through electrophilic alkylation at its carbon-carbon double bond (18,19). The reaction depends for activation on a trinuclear metal cluster, usually containing Mg 2+ or Mn 2+ (20). Based on earlier studies describing the role of metal cofactors for scIDS catalysis, we tested the product composition of a unique scIDS discovered from juvenile horseradish leaf beetles, Phaedon cochleariae, in the presence of different metal ions.…”

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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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Trinuclear metal clusters in catalysis by terpenoid synthases

Cited by 127 publications

References 57 publications

Structure and Function of Fusicoccadiene Synthase, a Hexameric Bifunctional Diterpene Synthase

Structure and Function of Fusicoccadiene Synthase, a Hexameric Bifunctional Diterpene Synthase

Molecular Diversity of Terpene Synthases in the Liverwort Marchantia polymorpha

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

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