Structure and Function of Fusicoccadiene Synthase, a Hexameric Bifunctional Diterpene Synthase
Abstract
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.
- Authors:
-
- 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)
- Publication Date:
- Research Org.:
- Argonne National Laboratory (ANL), Argonne, IL (United States)
- Sponsoring Org.:
- USDOE; National Institutes of Health (NIH)
- OSTI Identifier:
- 1249236
- Grant/Contract Number:
- AC02-06CH11357; P41 GM103403; S10 RR029205
- Resource Type:
- Accepted Manuscript
- Journal Name:
- ACS Chemical Biology
- Additional Journal Information:
- Journal Volume: 11; Journal Issue: 4; Journal ID: ISSN 1554-8929
- Publisher:
- American Chemical Society (ACS)
- Country of Publication:
- United States
- Language:
- ENGLISH
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; 37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY
Citation Formats
Chen, Mengbin, Chou, Wayne K. W., Toyomasu, Tomonobu, Cane, David E., and Christianson, David W. Structure and Function of Fusicoccadiene Synthase, a Hexameric Bifunctional Diterpene Synthase. United States: N. p., 2016.
Web. doi:10.1021/acschembio.5b00960.
Chen, Mengbin, Chou, Wayne K. W., Toyomasu, Tomonobu, Cane, David E., & Christianson, David W. Structure and Function of Fusicoccadiene Synthase, a Hexameric Bifunctional Diterpene Synthase. United States. https://doi.org/10.1021/acschembio.5b00960
Chen, Mengbin, Chou, Wayne K. W., Toyomasu, Tomonobu, Cane, David E., and Christianson, David W. Wed .
"Structure and Function of Fusicoccadiene Synthase, a Hexameric Bifunctional Diterpene Synthase". United States. https://doi.org/10.1021/acschembio.5b00960. https://www.osti.gov/servlets/purl/1249236.
@article{osti_1249236,
title = {Structure and Function of Fusicoccadiene Synthase, a Hexameric Bifunctional Diterpene Synthase},
author = {Chen, Mengbin and Chou, Wayne K. W. and Toyomasu, Tomonobu and Cane, David E. and Christianson, David W.},
abstractNote = {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.},
doi = {10.1021/acschembio.5b00960},
journal = {ACS Chemical Biology},
number = 4,
volume = 11,
place = {United States},
year = {Wed Jan 06 00:00:00 EST 2016},
month = {Wed Jan 06 00:00:00 EST 2016}
}
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