Investigating substrate channeling and cooperativity in assembly-line terpenoid synthases.
EMSL Project ID
50842
Abstract
Bifunctional assembly-line terpenoid synthases catalyze a specific sequence of chemical reactions starting with simple 5-carbon precursors, and ending with 20- or 25-carbon polycyclic natural products that contain multiple fused rings and stereocenters. Many such terpenoids exhibit useful biological properties, e.g., as anti-cancer, anti-inflammatory, or antibacterial agents. Terpenoid synthase reaction sequences are among the most complex carbon-carbon bond forming reactions in nature, since approximately two-thirds of the carbon atoms in the substrates undergo changes in bonding and/or hybridization during a single, precisely controlled reaction sequence. Recent discoveries reveal oligomeric terpenoid synthases that catalyze consecutive reaction in an assembly-line fashion. Although we have determined crystal structures of individual catalytic domains for one assembly-line synthase, the structure of an intact oligomeric synthases is yet unknown and refractory to crystallization. Only the structure of an intact oligomer will reveal the structural basis of substrate channeling between active sites and the molecular basis of cooperativity measured in enzymological studies. Cryo-EM is thus the best approach for structure determination of intact oligomers at atomic resolution. The studies proposed herein will directly inform biosynthesis of pharmaceutically relevant terpenoids.
Project Details
Start Date
2019-05-15
End Date
2019-07-10
Status
Closed
Released Data Link
Team
Principal Investigator
Team Members
Related Publications
Faylo, Jacque L., et al. "Structural insight on assembly-line catalysis in terpene biosynthesis." Nature Communications 12.1 (2021): 1-12.