In the area of terpenoid biosynthesis, Professor Cane's group is isolating, identifying, purifying and characterizing a variety of new enzymes which catalyze the conversion of the universal acyclic precursors, farnesyl and geranylgeranyl diphosphate, to cyclic sesquiterpenes and diterpenes, respectively. In the course of this work they have established many key details of this fascinating cyclization process and developed both reversible and irreversible inhibitors of the cyclases. In addition, cloning, sequencing and overexpression of terpenoid synthase genes has provided large quantities of protein for further mechanistic and protein structural studies. Site directed mutagenesis experiments are shining light on the mechanisms of cylclization of the universal precursors.

For example, it has been found that a series of trichodiene synthase mutants generated by site-directed mutagensis produce of family of anomalous cyclization products, all of which appear to result from premature quenching of the otherwise cryptic intermediates of the normal cyclization. Recently, in collaboration with Prof. David Christianson of the University of Pennsylvania, the crystal structure of pentalenene synthase and aristolochene synthase from Penicillium roqueforti have been solved.

Representative Publications

Michael J. Rynkiewicz, David E. Cane, and David W. Christianson, "Structure of trichodiene synthase from Fusarium sporotrichioides provides mechanistic inferences on the terpene cyclization cascade," Proc. Natl. Acad. Sci. USA 98, 13543-13548 (2001).

David E. Cane and Ilgu Kang, "Aristolochene Synthase. Purification, Molecular Cloning, High-Level Expression in Escherichia coli, and Characterization of the Aspergillus terreus Cyclase," Arch. Biochem. Biophys. 376, 354-364 (2000).

Jonathan M. Caruthers, Ilgu Kang, Michael J. Rynkiewicz, David E. Cane, and David W. Christianson, "Crystal Structure Determination of Aristolochene Synthase from the Blue Cheese Mold, Penicillium roqueforti,", J. Biol. Chem. 275, 25533-25539 (2000).

David E. Cane and Todd E. Bowser, "Trichodiene Synthase. Mechanism-Based Inhibition Of A Sesquiterpene Cyclase," Bioorg. Med. Chem. Lett. 9, 1127-1132 (1999).

Myriam Seemann, Guangzhi Zhai, Kazuo Umezawa and David Cane, "Pentalenene Synthase. Histidine-309 is Not Required for Catalytic Activity," J. Am. Chem. Soc. 121, 591-592 (1999).

Charles A. Lesburg, Guangzhi Zhai, David E. Cane, and David W. Christianson, "Crystal Structure of Pentalenene Synthase: Mechanistic Insights on Terpenoid Cyclization Reactions in Biology," Science, 277, 1820-1824 (1997).

David E. Cane, Hsien-Tai Chiu, Po-Huang Liang, and Karen S. Anderson, "A Pre-Steady State Kinetic Analysis of the Trichodiene Synthase Reaction Pathway," Biochemistry, 36 , 8332 -8339 (1997).

David E. Cane, Qun Xue, and Brian C. Fitzsimons, "Trichodiene Synthase. Probing the Role of the Highly Conserved Aspartate-Rich Region by Site-Directed Mutagenesis," Biochemistry, 35, 12369-12376 (1996).

David E. Cane, Qun Xue, and James E. Van Epp, "Enzymatic Formation of Isochamigrene, a Novel Sesquiterpene, by Alteration of the Aspartate-Rich Region of Trichodiene Synthase," J. Am. Chem. Soc. 118, 8499-8500 (1996).

David E. Cane and Youla S. Tsantrizos, "Aristolochene Synthase. Elucidation of the Cryptic Germacrene A Synthase Activity Using the Anomalous Substrate Dihydrofarnesyl Diphosphate," J. Am. Chem. Soc. 118, 10037-10040 (1996).

Alison Hill, David E. Cane, Christopher D. Mau, and Charles West, "High Level Expression of Ricinus communis Casbene Synthase in Escherichia coli and Characterization of the Recombinant Enzyme," Arch. Biochem. Biophys. 336, 283-289 (1996).

David E. Cane and Youla S. Tsantrizos, "Aristolochene Synthase. Elucidation of the Cryptic Germacrene A Synthase Activity Using the Anomalous Substrate Dihydrofarnesyl Diphosphate," J. Am. Chem. Soc. 118, 10037-10040 (1996).

David E. Cane, Qun Xue, and James E. Van Epp, "Enzymatic Formation of Isochamigrene, a Novel Sesquiterpene, by Alteration of the Aspartate-Rich Region of Trichodiene Synthase," J. Am. Chem. Soc. 118, 8499-8500 (1996).

David E. Cane, Qun Xue, and Brian C. Fitzsimons, "Trichodiene Synthase. Probing the Role of the Highly Conserved Aspartate-Rich Region by Site-Directed Mutagenesis," Biochemistry, 35, 12369-12376 (1996).

David E. Cane and Qun Xue, "Trichodiene Synthase. Enzymatic Formation of Multiple Sesquiterpenes by Alteration of the Cyclase Active Site," J. Am. Chem. Soc. , 118, 1563-1564 (1996)