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| Title | Transient Prenyltransferase-Cyclase Association in Fusicoccadiene Synthase, an Assembly-Line Terpene Synthase. |
|---|---|
| Journal, issue, pages | Biochemistry, Vol. 61, Issue 21, Page 2417-2430, Year 2022 |
| Publish date | Nov 1, 2022 |
 Authors | Jacque L Faylo / Trevor van Eeuwen / Kushol Gupta / Kenji Murakami / David W Christianson /  |
| PubMed Abstract | Fusicoccadiene synthase from the fungus (PaFS) is an assembly-line terpene synthase that catalyzes the first two steps in the biosynthesis of Fusiccocin A, a diterpene glycoside. The C-terminal ...Fusicoccadiene synthase from the fungus (PaFS) is an assembly-line terpene synthase that catalyzes the first two steps in the biosynthesis of Fusiccocin A, a diterpene glycoside. The C-terminal prenyltransferase domain of PaFS catalyzes the condensation of one molecule of C dimethylallyl diphosphate and three molecules of C isopentenyl diphosphate to form C geranylgeranyl diphosphate, which then transits to the cyclase domain for cyclization to form fusicoccadiene. Previous structural studies of PaFS using electron microscopy (EM) revealed a central octameric prenyltransferase core with eight cyclase domains tethered in random distal positions through flexible 70-residue linkers. However, proximal prenyltransferase-cyclase configurations could be captured by covalent cross-linking and observed by cryo-EM and mass spectrometry. Here, we use cryo-EM to show that proximally configured prenyltransferase-cyclase complexes are observable even in the absence of covalent cross-linking; moreover, such complexes can involve multiple cyclase domains. A conserved basic patch on the prenyltransferase domain comprises the primary touchpoint with the cyclase domain. These results support a model for transient prenyltransferase-cyclase association in which the cyclase domains of PaFS are in facile equilibrium between proximal associated and random distal positions relative to the central prenyltransferase octamer. The results of biophysical measurements using small-angle X-ray scattering, analytical ultracentrifugation, dynamic light scattering, and size-exclusion chromatography in-line with multi-angle light scattering are consistent with this model. This model accordingly provides a framework for understanding substrate transit between the prenyltransferase and cyclase domains as well as the cooperativity observed for geranylgeranyl diphosphate cyclization. |
 External links | Biochemistry / PubMed:36227241 / PubMed Central |
| Methods | EM (single particle) |
| Resolution | 3.73 - 8.4 Å |
| Structure data | EMDB-27989, PDB-8eax:  EMDB-28046: Symmetry-expanded consensus map of PaFS prenyltransferase core with single interacting cyclase domain  EMDB-28076: Symmetry-expanded map of PaFS prenyltransferase core with single interacting cyclase domain class A  EMDB-28079: Symmetry-expanded map of PaFS prenyltransferase core with single interacting cyclase domain, class B  EMDB-28131: PaFS prenyltransferase core with single interacting cyclase domain - North class A  EMDB-28132: PaFS prenyltransferase core with single interacting cyclase domain - North class D  EMDB-28191: PaFS prenyltransferase core with single interacting cyclase domain - North class C  EMDB-28193: PaFS prenyltransferase core with single interacting cyclase domain - North class B  EMDB-28194: PaFS prenyltransferase core with single interacting cyclase domain - East class D  EMDB-28201: PaFS prenyltransferase core with single interacting cyclase domain - East class B  EMDB-28202: PaFS prenyltransferase core with single interacting cyclase domain - East class A  EMDB-28203: PaFS prenyltransferase core with single interacting cyclase domain - East class C  EMDB-28213: PaFS prenyltransferase core with single interacting cyclase domain - South class A  EMDB-28215: PaFS prenyltransferase core with single interacting cyclase domain - South class B  EMDB-28216: PaFS prenyltransferase core with single interacting cyclase domain - South class C  EMDB-28219: PaFS prenyltransferase core with single interacting cyclase domain - West class A  EMDB-28220: PaFS prenyltransferase core with single interacting cyclase domain - West class B  EMDB-28221: PaFS prenyltransferase core with single interacting cyclase domain - West class C  EMDB-28222: PaFS prenyltransferase core with single interacting cyclase domain - West class D  EMDB-28235: PaFS prenyltransferase core with two interacting cyclase domains - East and South  EMDB-28236: PaFS prenyltransferase core with two interacting cyclase domains - South and East  EMDB-28237: PaFS prenyltransferase core with two interacting cyclase domains - East and West  EMDB-28239: PaFS prenyltransferase core with two interacting cyclase domains - West and East  EMDB-28240: PaFS prenyltransferase core with two interacting cyclase domains - North and East  EMDB-28245: PaFS prenyltransferase core with two interacting cyclase domains - East and North |
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 Keywords | TRANSFERASE / Terpene synthase / Prenyltransferase |
diaporthe amygdali (fungus)
Escherichia coli (E. coli)