Studies on the nonmevalonate pathway: conversion of 4-(cytidine 5′-diphospho)-2-C-methyl-d-erythritol to its 2-phospho derivative by 4-(cytidine 5′-diphospho)-2-C-methyl-d-erythritol kinase

Kuzuyama, Tomohisa; Takagi, Motoki; Kaneda, Kazuhide; Watanabe, Hiroyuki; Dairi, Tohru; Seto, Haruo

doi:10.1016/s0040-4039(00)00295-1

Cited by 105 publications

(66 citation statements)

References 4 publications

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“…Compound 3 is converted into the branched chain polyol 2C-methyl-D-erythritol 4-phosphate 4 by skeletal rearrangement and reduction (11). The polyol 4 is converted via 4-diphosphocytidyl-2C-methyl-D-erythritol 5 and its 2-phosphate 6 into 2C-methyl-D-erythritol 2,4-cyclodiphosphate 7 by a sequence of three reaction steps catalyzed by proteins that are specified by the ispDEF genes (12)(13)(14)(15)(16)(17). In vivo experiments showed that the reductive ring opening of 7 affording 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate 8 requires a protein specified by the ispG (gcpE) gene (18,19).…”

Section: Earlier In Vivo Studies Showed the Involvement Of Isph Protementioning

confidence: 99%

Biosynthesis of terpenes: Studies on 1-hydroxy-2-methyl-2-( E )-butenyl 4-diphosphate reductase

Adam

Hecht²,

Eisenreich³

et al. 2002

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

152

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Earlier in vivo studies showed the involvement of IspH protein in the conversion of 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate into isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP). We have demonstrated now that cell extract of an Escherichia coli strain engineered for hyperexpression of the ispH (lytB) gene catalyzes the in vitro conversion of 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate into IPP and DMAPP. The reaction requires NADH, FAD, divalent cations (preferably Co 2؉ ), and probably one or more as-yet-unidentified proteins. The low intrinsic catalytic activities of wild-type E. coli cell extract and isolated chromoplasts of red pepper (Capsicum annuum) are enhanced by the addition of purified recombinant IspH protein.

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Section: Earlier In Vivo Studies Showed the Involvement Of Isph Protementioning

confidence: 99%

Biosynthesis of terpenes: Studies on 1-hydroxy-2-methyl-2-( E )-butenyl 4-diphosphate reductase

Adam

Hecht²,

Eisenreich³

et al. 2002

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

152

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“…7 This reaction is catalyzed by NADPH-dependent DXP reductoisomerase. 8 MEP is finally converted into both IPP and dimethylallyl diphosphate by the action of the following five enzymes, MEP cytidylyltransferase, 9,10 4-(cytidine 5¢-diphospho)-2-C-methyl-D-erythritol kinase, 11,12 2-C-methyl-D-erythritol 2,4-cyclodiphosphate synthase, 13,14 4-hydroxy-3-methylbut-2-en-1-yl diphosphate synthase 15 and 4-hydroxy-3-methylbut-2-en-1-yl diphosphate reductase. 16 The MEP pathway is present in most bacteria, including pathogens, as well as in plant chloroplasts, and the apicoplasts of Apicomplexan protozoa such as Plasmodium and Toxoplasma.…”

Section: Introductionmentioning

confidence: 99%

The herbicide ketoclomazone inhibits 1-deoxy-D-xylulose 5-phosphate synthase in the 2-C-methyl-D-erythritol 4-phosphate pathway and shows antibacterial activity against Haemophilus influenzae

et al. 2010

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Two distinct metabolic pathways have been elucidated for the formation of isopentenyl diphosphate and dimethylallyl diphosphate, essential metabolic precursors for isoprenoid biosynthesis: the mevalonate pathway, found ubiquitously in mammals, and the 2-C-methyl-D-erythritol 4-phosphate (MEP) pathway, found in most bacteria. As the MEP pathway is absent from mammals, all MEP pathway enzymes represent effective targets for the development of antibacterial drugs. In this study, we found that a herbicide, ketoclomazone, exhibited antibacterial activity against a pathogenic bacterium, Haemophilus influenzae, with an MIC value of 12.5 lg ml À1 and that antibacterial activity was suppressed by adding 1-deoxyxylulose, a free alcohol of 1-deoxy-D-xylulose 5-phosphate (DXP). DXP is an MEP pathway intermediate synthesized from pyruvate and D-glyceraldehyde 3-phosphate (D-GAP) by the action of DXP synthase. Thus, we investigated the enzyme kinetics of DXP synthase of H. influenzae (HiDXS) to elucidate an inhibitory mechanism of ketoclomazone on HiDXS. The dxs gene was cloned from H. influenzae and overexpressed in Escherichia coli, and the enzyme was purified to homogeneity. The purified HiDXS was a soluble dimeric 70-kDa protein. Steady-state kinetic constants for HiDXS were calculated, and Lineweaver-Burk plots were consistent with a ping-pong bi bi mechanism. The kinetics of inhibition by ketoclomazone suggested that ketoclomazone binds to an unidentified inhibitor-binding site that differs from both the pyruvate-binding site and the D-GAP-binding site on DXP synthase. These data reveal the inhibitory mechanism of ketoclomazone on DXP synthase.

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“…The second and third stages convert DOXP to 2C-methyl-D-erythritol-4-phosphate (11, 12) and transfer the erythritol derivative onto a nucleotide, resulting in CDP-ME (13-15). Stage four, the only ATP-dependent step in the pathway, is catalyzed by CDP-ME kinase (16,17), the subject of the present study. Here, CDP-ME kinase catalyzes the transfer of the ␥-phosphoryl moiety of ATP to CDP-ME, forming 4-diphosphocytidyl-2C-methyl-D-erythritol-2-phosphate (CDP-ME2P) and ADP (Fig.…”

mentioning

confidence: 96%

Biosynthesis of isoprenoids: Crystal structure of 4-diphosphocytidyl-2C-methyl- d -erythritol kinase

Miallau

Alphey

Kemp

et al. 2003

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

109

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4-Diphosphocytidyl-2C-methyl-D-erythritol kinase, an essential enzyme in the nonmevalonate pathway of isopentenyl diphosphate and dimethylallyl diphosphate biosynthesis, catalyzes the single ATP-dependent phosphorylation stage affording 4-diphosphocytidyl-2C-methyl-D-erythritol-2-phosphate. The 2-Å resolution crystal structure of the Escherichia coli enzyme in a ternary complex with substrate and a nonhydrolyzable ATP analogue reveals the molecular determinants of specificity and catalysis. The enzyme subunit displays the ␣͞␤ fold characteristic of the galactose kinase͞ homoserine kinase͞mevalonate kinase͞phosphomevalonate kinase superfamily, arranged into cofactor and substrate-binding domains with the catalytic center positioned in a deep cleft between domains. Comparisons with related members of this superfamily indicate that the core regions of each domain are conserved, whereas there are significant differences in the substrate-binding pockets. The nonmevalonate pathway is essential in many microbial pathogens and distinct from the mevalonate pathway used by mammals. The high degree of sequence conservation of the enzyme across bacterial species suggests similarities in structure, specificity, and mechanism. Our model therefore provides an accurate template to facilitate the structure-based design of broad-spectrum antimicrobial agents.galactose kinase͞homoserine kinase͞mevalonate kinase͞ phosphomevalonate kinase ͉ enzyme mechanism ͉ nonmevalonate ͉ phosphorylation T he AT P-dependent 4-diphosphocytidyl-2C-methyl-Derythritol (CDP-ME) kinase (EC 2.7.1.148) participates in the biosynthesis of isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP). These isomers are the universal five-carbon precursors of isoprenoids, a diverse and important family of natural products that includes sterols, dolichols, triterpenes, and ubiquinones, and components of macromolecules such as the prenyl groups of prenylated proteins and isopentenylated tRNAs (1-3). Isoprenoids contribute to many biological functions, including electron transport in respiration and photosynthesis, hormone-based signaling, apoptosis, meiosis, protein cleavage, and degradation (4). In addition, they provide important structural components of cell membranes (1).Two biosynthetic routes to IPP and DMAPP have evolved. In eukaryotes, archaebacteria, and a few eubacteria, the precursor biosynthesis is through the mevalonate pathway (3-7). This begins with the conversion of three molecules of acetyl-CoA to 3-hydroxy-3-methylglutaryl-CoA followed by reduction, phosphorylation, and decarboxylation to generate IPP, some of which is isomerized to DMAPP. The last three steps in this pathway are ATP-dependent and catalyzed by the structurally related mevalonate kinase (MVK), phosphomevalonate kinase, and mevalonate 5-diphosphate decarboxylase.In chloroplasts, algae, cyanobacteria, most eubacteria, and the apicomplexa, IPP and DMAPP synthesis is accomplished by seven enzymes in a pathway named after one of the intermediates, the 1-deoxy-D-xylulose-5-ph...

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Studies on the nonmevalonate pathway: conversion of 4-(cytidine 5′-diphospho)-2-C-methyl-d-erythritol to its 2-phospho derivative by 4-(cytidine 5′-diphospho)-2-C-methyl-d-erythritol kinase

Cited by 105 publications

References 4 publications

Biosynthesis of terpenes: Studies on 1-hydroxy-2-methyl-2-( E )-butenyl 4-diphosphate reductase

Biosynthesis of terpenes: Studies on 1-hydroxy-2-methyl-2-( E )-butenyl 4-diphosphate reductase

The herbicide ketoclomazone inhibits 1-deoxy-D-xylulose 5-phosphate synthase in the 2-C-methyl-D-erythritol 4-phosphate pathway and shows antibacterial activity against Haemophilus influenzae

Biosynthesis of isoprenoids: Crystal structure of 4-diphosphocytidyl-2C-methyl- d -erythritol kinase

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