Distinguished Professor of Chemistry and
Adjunct Professor of Biochemistry
B.S. Louisiana State University
Ph.D. University of California, Berkeley
Dale Poulter's Lab Page
Dale Poulter's PubMed Literature Search
Biological Chemistry Program
Biosynthesis, Structure Function, Mechanistic Enzymology
My research group studies how biosynthetic pathways emerge through the evolution of new enzyme activities. We use a variety of biochemical, microbiological, and molecular biological techniques to determine structure-function relationships, with a special emphasis on understanding how the active sites of enzymes can be molded to efficiently synthesize new compounds. Our interests include enzymes in the isoprene synthase superfamily, which are responsible for synthesis of over 60,000 naturally occurring molecules, the independently evolved mevalonate and methylerythritol phosphate pathways for synthesis of the isoprenoid building blocks, and the enzymes that attach isoprenoid units to non-isoprenoid acceptors in the posttranscriptional modification of tRNA, the posttranslational modification of proteins, and the biosynthesis of amino acid and quinoid metabolites.
Enzymes in the isoprene synthase superfamily catalyze the eight basic bond-forming reactions to construct the carbon skeletons of metabolites in the isoprenoid pathway, each with high selectivity for a specific reaction. These molecules are essential for life and the pathway is found in all organisms. Our structure-function studies show that the high selectivity of isoprene synthases can be altered by mutagenesis to give enzymes that produce products with all but one of the carbon skeletons formed in nature and provide insight about how new enzyme activities can be developed from a parental protein scaffold.
Eukaryota and Archaea synthesize isoprenoid molecules from acetyl CoA by the mevalonate pathway. However the genomes of Archaea lack two of the enzymes found in eukaryotes. We have characterized the kinetic properties and determined the X-ray structure of one of the missing enzymes and are working to discover the missing the gene for the missing enzyme. Our work has led to the discovery new biosynthetic enzymes and an understanding of how they are related to other members of the isoprenoid synthase superfamily. We are currently using genome screening, biochemical characterization and structural methods to understand the evolutionary relationships in this group of enzymes.
- Choi, S.-R..; Seo, J.-S.; Bohaty, R. F. H.; Poulter, C.D. “Regio- and Chemoselective Immobilization of Proteins on Gold Surfaces” Bioconjugate Chem. 2014, 25, 269-275.
- de Ruyck, J.; Janczak, M. W.; Neti, S. S.; Rothman, S. C.; Schubert, H. L; Cornish, R. M.; Matagne, A.; Wouters, J.; Poulter, C. D. “Determination of Kinetics and the Crystal Structure of a Novel Type 2 Isopentenyl Diphosphate: Dimethylallyl Diphosphate Isomerase from Streptococcus pneumonia” ChemBioChem, 2014, 15, 1452-1458.
- Krasutsky, S.G.; Urbansky, M.; Davis, C.E.; Lherbet, C.; Coates, R.M.; Poulter, C.D. “Synthesis of Methylerythritol Phosphate Analogues and their Evaluation as Alternate Substrates for IspDF and IspE from Agrobacterium tumefaciens” J. Org. Chem., 2014, 79, 9170-9178.
- Tian, B.; Wallrapp, F.H.; Holiday, G.L.; Chow, J.-Y.; Babbitt, P.C.; Poulter, C.D.; Jacobson, M.P. “Predicting the Functions and Specificity of Triterpenoid Synthases: A Mechanism-Based Multi-Intermediate Docking Approach” PLoS Computational Biology, 2014, 10, e1003874.
- Pan, J.-J.; Solbiati, J.O.; Ramamoorthy, G.; Hillerich, B. S.; Seidel, R. D.; Cronan, J. E.; Almo, S. C.; Poulter, C. D. “Biosynthesis of Squalene from Farnesyl Diphosphate in Bacteria: Three Steps Catalyzed by Three Enzymes” ACS Cent. Sci., 2015, 1, 77-82.
- Chow, J.-Y.; Tian, B.-X.; Ramamoorthy, G.; Hillerich, B. S.; Seidel, R. D.; Almo, S. C.; Jacobson, M. P.; Poulter, C. D. “Computational-guided discovery and characterization of a sesquiterpene synthase from Streptomyces clavuligerus” Proc. Nat. Acad. Sci. USA, 2015, 112, 5661-5666.
- Ramamoorthy, G.; Mark, L. P.; Tian, B.-X.; Phan, R.M.; Perez, L. B.; Jacobson, M.P.; Poulter, C. D. “Synthesis and enzymatic studies of bisubstrate analogues for farnesyl diphosphate synthase” J. Org. Chem, 2015, 80, 3902-3913.
- Faus, I.; Reinhard, A.; Rackwitz, S.; Wolny, J. A.; Schlage, K.; Wille, H.-C.; Chumakov, A.; Krasutsky, S.; Chaignon, P.; Poulter, C. D.; Seeman, M.; Schunemann, V. “Isoprenoid Biosynthesis in Pathogenic Bacteria: Nuclear Resonance Vibrational Spectroscopy Provides Insight into the Unusual [4Fe-4S] Cluster of the E. coli LytB/IspH Protein” Angew. Chem. Int. Ed., 2015, 54, 12584-12587.
- Pan, J.-J.; Ramamoorthy, G.; Poulter, C. D. “Absolute Configuration of Hydroxysqualene. An Intermediate in Bacterial Hopanoid Biosynthesis” Org. Lett., 2016, 18, 512-515.
- Tian, B.; Poulter, C.D.; Jacobson, M.P. “Defining the Product Chemical Space of Monoterpenoid Synthases” PLOS Comput Biol, 2016, 12, e1005053.
- Neti, S.S.; Poulter, C.D. “Site-Selective Synthesis of 15N- and 13C‑Enriched Flavin Mononucleotide Coenzyme Isotopologues” J. Org. Chem.,2016, 81, 5087−5092.
- Ramamoorthy, G.; Phan, R.M.; Poulter, C.D. “Synthesis and Enzymatic Studies of Isoprenoid Thiolo Bisubstrate Analogues” J. Org. Chem.,2016, 81, 5093−5100.
- Neti, S.S.; Eckertand, D.M.; Poulter, C.D. “Construction of Functional Monomeric Type 2 Isopentenyl Diphosphate:Dimethylallyl Diphosphate Isomerase” Biochemistry, 2016, 55, 4229-4238.
- Janczak, M.W.; Poulter, C.D. “Kinetic and Binding Studies of Streptococcus pneumoniae Type 2 Isopentenyl Diphosphate:Dimethylallyl Diphosphate Isomerase” Biochemistry, 2016, 55, 2260−2268.