Hedycaryol Synthase in Complex with Nerolidol Reveals Terpene Cyclase Mechanism

Baer, Philipp; Rabe, Patrick; Citron, Christian A.; Mann, Carina C. de Oliveira; Kaufmann, N.; Groll, M.; Dickschat, Jeroen S.

doi:10.1002/cbic.201300708

Cited by 70 publications

(115 citation statements)

References 27 publications

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“…The best fit of fusicoccadiene in the active site is such that sites of developing positive charge on the face of the macrocycle are oriented toward the backbone carbonyl of V192, consistent with possible carbocation stabilization by the helix G1 macrodipole. 39 …”

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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“…Crystal structures have been solved for several microbial and plant sesquiterpene synthases [83,84,[115][116][117][118][119][120][121]. Aristolochene synthase from Penicillium roqueforti and Aspergillus terreus [84,117,122,123] and trichodiene synthase from Fusarium sporotrichioides [83,103,124,125] The active site of all sesquiterpene synthases is located in a α-helical bundle (α-domain), which is characteristic for ionization-dependent class I terpene synthases as discussed above.…”

Section: Overviewmentioning

confidence: 99%

Biosynthesis of Terpenoid Natural Products in Fungi

Schmidt‐Dannert

2014

Advances in Biochemical Engineering/Biotechnology

108

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Tens of thousands of terpenoid natural products have been isolated from plants and microbial sources. Higher fungi (Ascomycota and Basidiomycota) are known to produce an array of well-known terpenoid natural products, including mycotoxins, antibiotics, antitumor compounds, and phytohormones. Except for a few well-studied fungal biosynthetic pathways, the majority of genes and biosynthetic pathways responsible for the biosynthesis of a small number of these secondary metabolites have only been discovered and characterized in the past 5-10 years. This chapter provides a comprehensive overview of the current knowledge on fungal terpenoid biosynthesis from biochemical, genetic, and genomic viewpoints. Enzymes involved in synthesizing, transferring, and cyclizing the prenyl chains that form the hydrocarbon scaffolds of fungal terpenoid natural products are systematically discussed. Genomic information and functional evidence suggest differences between the terpenome of the two major fungal phyla--the Ascomycota and Basidiomycota--which will be illustrated for each group of terpenoid natural products.

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“…62 Pentalenene synthase, 40 while having a slightly lower sequence identity (26%) with the N-terminal domain of ScGS, has a higher sequence similarity (41%). ScGS 338 has nearly identical helix topology to that found in the structures of pentalenene synthase and epi -isozizaene synthase (EIZS).…”

Section: Resultsmentioning

confidence: 99%

“…The active site contours of enzymes that direct initial 1,6-cyclizations, such as epi -isozizaene synthase 60 and trichodiene synthase, 73 tend to be somewhat narrow and deep. In contrast, enzymes that direct 1,10-cyclization reactions, such as selinadiene synthase 58 (via direct formation of a ( E , E )-germacradienyl cation intermediate) and hedycaryol synthase 62 (via initial isomerization to nerolidyl diphosphate followed by formation of a ( Z , E )-helminthogermacradienyl cation), as well as an enzyme that directs a 1,11-cyclization reaction, pentalenene synthase, 40 all appear to have wider, shallower active sites (Figure S3, Supporting Information). …”

Section: Discussionmentioning

confidence: 99%

Structural Studies of Geosmin Synthase, a Bifunctional Sesquiterpene Synthase with αα Domain Architecture That Catalyzes a Unique Cyclization–Fragmentation Reaction Sequence

et al. 2015

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Geosmin synthase from Streptomyces coelicolor (ScGS) catalyzes an unusual, metal-dependent terpenoid cyclization and fragmentation reaction sequence. Two distinct active sites are required for catalysis: the N-terminal domain catalyzes the ionization and cyclization of farnesyl diphosphate to form germacradienol and inorganic pyrophosphate (PPi), and the C-terminal domain catalyzes the protonation, cyclization, and fragmentation of germacradienol to form geosmin and acetone through a retro-Prins reaction. A unique αα domain architecture is predicted for ScGS based on amino acid sequence: each domain contains the metal-binding motifs typical of a class I terpenoid cyclase, and each domain requires Mg2+ for catalysis. Here, we report the X-ray crystal structure of the unliganded N-terminal domain of ScGS and the structure of its complex with 3 Mg2+ ions and alendronate. These structures highlight conformational changes required for active site closure and catalysis. Although neither full-length ScGS nor constructs of the C-terminal domain could be crystallized, homology models of the C-terminal domain were constructed based on ~36% sequence identity with the N-terminal domain. Small-angle X-ray scattering experiments yield low resolution molecular envelopes into which the N-terminal domain crystal structure and the C-terminal domain homology model were fit, suggesting possible αα domain architectures as frameworks for bifunctional catalysis.

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Hedycaryol Synthase in Complex with Nerolidol Reveals Terpene Cyclase Mechanism

Cited by 70 publications

References 27 publications

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

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

Biosynthesis of Terpenoid Natural Products in Fungi

Structural Studies of Geosmin Synthase, a Bifunctional Sesquiterpene Synthase with αα Domain Architecture That Catalyzes a Unique Cyclization–Fragmentation Reaction Sequence

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