Model studies of the biosynthesis of non-head-to-tail terpenes. Rearrangements of the chrysanthemyl system

Poulter, C. Dale; Marsh, Larry L.; Hughes, J.; Argyle, J. Craig; Satterwhite, D M; Goodfellow, Robyn J.; Moesinger, S. G.

doi:10.1021/ja00453a050

Cited by 42 publications

(37 citation statements)

References 11 publications

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“…There are eight different ways in which the individual isoprene units are attached by these building reactions (see Figure 7). Our model studies of the chrysanthemyl cation suggested that four of the structures, the 1′-1, 1′-3, c1′-1-2, and 2-1′-3 skeletons, are derived from the c1′-2-3 structure by rearrangement 16–18. The 1′-4 skeleton is produced by condensation of an allylic diphosphate with isopentenyl diphosphate 45.…”

Section: Studies With Recombinant Enzymesmentioning

confidence: 86%

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Bioorganic Chemistry. A Natural Reunion of the Physical and Life Sciences

Poulter

2009

J. Org. Chem.

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Organic substances were conceived as those found in living organisms. Although the definition was soon broadened to include all carbon-containing compounds, naturally occurring molecules have always held a special fascination for organic chemists. From these beginnings, molecules from nature were indespensible tools as generations of organic chemists developed new techniques for determining structures, analyzed the mechanisms of reactions, explored the effects conformation and stereochemistry on reactions, and found challenging new targets to synthesize. Only recently have organic chemists harnessed the powerful techniques of organic chemistry to study the functions of organic molecules in their biological hosts, the enzymes that synthesize molecules and the complex processes that occur in a cell. In this Perspective, I present a personal account my entrée into bioorganic chemistry as a physical organic chemist and subsequent work to understand the chemical mechanisms of enzyme-catalyzed reactions, to develop techniques to identify and assign hydrogen bonds in tRNAs through NMR studies with isotopically labeled molecules, and to study how structure determines function in biosynthetic enzymes with proteins obtained by genetic engineering.As its name suggests, organic chemistry emerged in the early nineteenth century as a branch of chemistry concerned with substances isolated from living organisms. The field soon expanded, however, to include carbon-containing molecules more generally, and chemists began to study the structures, physical properties, reactions and chemical transformations of organic compounds, many of which were not obtained from nature. By the mid-twentieth century, most of the research in organic chemistry was not concerned with biological systems. Biology, on the other hand, focused largely on the morphologies and behaviors of organisms. Interpretation of biological phenomena at the molecular level was still in its infancy. Before that time, neither chemistry nor biology was sufficiently mature to nurture the other, and the research in each field was largely separate from the other. Thus, the great advances in both chemistry and biology before 1950 did not depend on insights from the other discipline.Today, however, the former artificial boundaries between organic chemistry and biology have been blurred as scientists in each area are quick to adopt the knowledge and techniques of the other. Natural products chemists analyze newly sequenced genomes for clues to previously undiscovered biologically active molecules and new biosynthetic pathways. Biologists identify molecules that regulate signaling events during cellular development and govern intereactions among and between species. Synthetic organic chemists rely on biological assays to guide their design of molecules that bind tightly to enzymes and receptors. And in my own research and that of others, the techniques of physical organic chemistry and molecular biology are applied to enzymes and other large biomolecules to gain an unders...

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Section: Studies With Recombinant Enzymesmentioning

confidence: 86%

“…The eventual answer was yes, but not very efficiently 16–18. When those experiments were carried out, there were no known natural monoterpenes with carbon skeletons corresponding to the tertiary cyclopropylcarbinyl cation and squalene, both of which we found in small amounts in the model studies (see Scheme 3).…”

Section: My Entrée Into Bioorganic Chemistrymentioning

confidence: 91%

Bioorganic Chemistry. A Natural Reunion of the Physical and Life Sciences

Poulter

2009

J. Org. Chem.

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“…Model studies subsequently demonstrated the feasibility of their proposal (2,28). As illustrated in Fig.…”

Section: Discussionmentioning

confidence: 87%

“…Chrysanthemol was not detected if the phosphatase step was omitted. Although a trace amount of yomogi alcohol (22) was detected in CPPase incubations longer than 1.5 h, control experiments indicated that the alcohol was from the nonenzymatic hydrolysis of CPP (28). Incubations of CPPase and radiolabeled IPP with DMAPP or geranyl diphosphate gave no evidence for formation of a 1Ј-4 product.…”

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confidence: 90%

Chrysanthemyl diphosphate synthase: Isolation of the gene and characterization of the recombinant non-head-to-tail monoterpene synthase from Chrysanthemum cinerariaefolium

Rivera

Swedlund

King

et al. 2001

Proc. Natl. Acad. Sci. U.S.A.

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Chrysanthemyl diphosphate synthase (CPPase) catalyzes the condensation of two molecules of dimethylallyl diphosphate to produce chrysanthemyl diphosphate (CPP), a monoterpene with a non-head-to-tail or irregular c1-2-3 linkage between isoprenoid units. Irregular monoterpenes are common in Chrysanthemum cinerariaefolium and related members of the Asteraceae family. In C. cinerariaefolium, CPP is an intermediate in the biosynthesis of the pyrethrin ester insecticides. CPPase was purified from immature chrysanthemum flowers, and the N terminus of the protein was sequenced. A C. cinerariaefolium cDNA library was screened by using degenerate oligonucleotide probes based on the amino acid sequence to identify a CPPase clone that encoded a 45-kDa preprotein. The first 50 aa of the ORF constitute a putative plastidial targeting sequence. Recombinant CPPase bearing an N-terminal polyhistidine affinity tag in place of the targeting sequence was purified to homogeneity from an overproducing Escherichia coli strain by Ni 2؉ chromatography. Incubation of recombinant CPPase with dimethylallyl diphosphate produced CPP. The diphosphate ester was hydrolyzed by alkaline phosphatase, and the resulting monoterpene alcohol was analyzed by GC͞MS to confirm its structure. The amino acid sequence of CPPase aligns closely with that of the chain elongation prenyltransferase farnesyl diphosphate synthase rather than squalene synthase or phytoene synthase, which catalyze c1-2-3 cyclopropanation reactions similar to the CPPase reaction.

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“…The cyclopropylcarbinyl pyrophosphates presqualene pyrophosphate and prephytoene pyrophosphate are essential intermediates in the biosynthesis of sterols (23) and carotenoids (24), respectively, whereas chrysanthemyl pyrophosphate, presumed to be synthesized by similar prenyl transfer, has been postulated as the key intermediate in the formation of irregular monoterpenes (25). Although the detailed mechanistic features of product formation from these cyclopropylcarbinyl pyrophosphates have not been unambiguously determined (23), and in spite of recent, alternate mechanistic rationales involving noncarbocationic processes (26,27), the prevailing opinion favors ionization to form a cyclopropylcarbinyl cationpyrophosphate anion pair that undergoes ring opening with concomitant proton loss (i.e., phytoene) or nucleophile capture (i.e., squalene and artetnisyl alcohol).…”

Section: Discussiotnmentioning

confidence: 99%