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

Chen, Mengbin; Chou, Wayne K. W.; Toyomasu, Tomonobu; Cane, David E.; Christianson, David W.

doi:10.1021/acschembio.5b00960

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

Journal Article · Wed Jan 06 00:00:00 EST 2016 · ACS Chemical Biology

DOI:https://doi.org/10.1021/acschembio.5b00960· OSTI ID:1249236

Chen, Mengbin ^[1]; Chou, Wayne K. W. ^[2]; Toyomasu, Tomonobu ^[3]; Cane, David E. ^[2]; Christianson, David W. ^[4]

Univ. of Pennsylvania, Philadelphia, PA (United States)
Brown Univ., Providence, RI (United States)
Yamagata Univ. (Japan)
Univ. of Pennsylvania, Philadelphia, PA (United States); Harvard Univ., Cambridge, MA (United States)

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 1930s. 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. Here, 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.

View Accepted Manuscript (DOE)

Research Organization:: Argonne National Laboratory (ANL), Argonne, IL (United States)

Sponsoring Organization:: NIH; USDOE

Grant/Contract Number:: AC02-06CH11357

OSTI ID:: 1249236

Journal Information:: ACS Chemical Biology, Journal Name: ACS Chemical Biology Journal Issue: 4 Vol. 11; ISSN 1554-8929

Publisher:: American Chemical Society (ACS)Copyright Statement

Country of Publication:: United States

Language:: ENGLISH

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