Expression and Characterization of Soluble 4-Diphosphocytidyl-2-C-methyl-D-erythritol Kinase from Bacterial Pathogens

Eoh, Hyungjin; Narayanasamy, Prabagaran; Brown, Alexander; Parish, Tanya; Brennan, Patrick J.; Crick, Dean C.

doi:10.1016/j.chembiol.2009.10.014

Cited by 23 publications

(26 citation statements)

References 34 publications

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“…We subsequently determined the K M for CDP-ME and ATP for the recombinant enzyme as 200μM and 20μM, respectively (data not shown). Our biochemical data correlated well with the data published by Rohdich and coworkers [32], as well as those of another recombinant bacterial CDP-ME kinase from Mycobacterium tuberculosis reported by Eoh and coworkers [33]. Therefore, His6 epitope tag did not appear to affect the overall folding of the E. coli enzyme and its function.…”

Section: Resultssupporting

confidence: 92%

Identification of novel small molecule inhibitors of 4-diphosphocytidyl-2-C-methyl-d-erythritol (CDP-ME) kinase of Gram-negative bacteria

Tang

Odejinmi

Allette

et al. 2011

Bioorganic & Medicinal Chemistry

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The biosyntheses of isoprenoids is essential for the survival in all living organisms, and requires one of the two biochemical pathways: (a) Mevalonate (MVA) Pathway or (b) Methylerythritol Phosphate (MEP) Pathway. The latter pathway, which is used by all Gram-negative bacteria, some Gram-positive bacteria and a few apicomplexan protozoa, provides an attractive target for the development of new antimicrobials because of its absence in humans. In this report, we describe two different approaches that we used to identify novel small molecule inhibitors of Escherichia coli and Yersinia pestis 4-diphosphocytidyl-2-C-methyl D-erythritol (CDP-ME) kinases, key enzymes of the MEP pathway encoded by the E. coli ispE and Y. pestis ipk genes, respectively. In the first approach, we explored existing inhibitors of the GHMP kinases while in the second approach; we performed computational high-throughput screening of compound libraries by targeting the CDP-ME binding site of the two bacterial enzymes. From the first approach, we identified two compounds with 6-(benzylthio)-2-(2-hydroxyphenyl)-4-oxo-3,4-dihydro-2H-1,3-thiazine-5-carbonitrile and (Z)-3-methyl-4-((5-phenylfuran-2-yl)methylene)isoxazol-5(4H)-one scaffolds which inhibited Escherichia coli CDP-ME kinase in vitro. We then performed substructure search and docking experiments based on these two scaffolds and identified twenty three analogs for structure-activity relationship (SAR) studies. Three new compounds from the isoxazol-5(4H)-one series have shown inhibitory activities against E. coli and Y. pestis CDP-ME kinases with the IC50 values ranging from 7μM to 13μM. The second approach by computational high-throughput screening (HTS) of two million drug-like compounds yielded two compounds with benzenesulfonamide and acetamide moieties which, at a concentration of 20μM, inhibited 80% and 65%, respectively, of control CDP-ME kinase activity.

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Section: Resultssupporting

confidence: 92%

Identification of novel small molecule inhibitors of 4-diphosphocytidyl-2-C-methyl-d-erythritol (CDP-ME) kinase of Gram-negative bacteria

Tang

Odejinmi

Allette

et al. 2011

Bioorganic & Medicinal Chemistry

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“…None of the reported structures of CMK indicate an associated metal ion, though a divalent metal ion was shown to be required for the activity of purified tomato CMK. 75 …”

Section: Generation Of C5 Isoprenoid Precursorsmentioning

confidence: 99%

Mechanistic Aspects of Carotenoid Biosynthesis

2013

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“…40–49 We preferred to synthesize 2- C -Methyl-D-erythritol-4-phosphoric acid ( 1 ) for CDP-ME synthesis instead of the its salts because as a free diacid, it could be used directly for CDP-ME synthesis 50 without the need for the extra steps for its salt preparation (sodium and trialkylammonium) and of the concerns for the limited solubility of the salts, particularly the sodium salt. 51 In our efforts to further simplify the synthesis of 2- C -Methyl-D-erythritol-4-phosphoric acid ( 1 ), we chose to explore its synthesis in 7 steps from D-(+)-arabitol ( 2 ) instead of the previously reported 14 steps.…”

Section: Resultsmentioning

confidence: 99%

Formal synthesis of 4-diphosphocytidyl-2-C-methyl d-erythritol from d-(+)-arabitol

et al. 2012

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2-C-methyl-D-erythritol-4-phosphate (MEP) is a key chemical intermediate of the non-mevalonate pathway for isoprenoid biosynthesis employed by many pathogenic microbes. MEP is also the precursor for the synthesis of 4-diphosphocytidyl-2-C-methyl D-erythritol (CDP-ME), another key intermediate of the non-mevalonate pathway. As this pathway is non-existent in higher animals, including humans, it represents great opportunities for novel antimicrobial development. To facilitate the in-depth studies of this pathway, we reported here a formal synthesis of CDP-ME through a new synthesis of 2-C-Methyl-D-erythritol-4-phosphoric acid from D-(+)-arabitol.

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Expression and Characterization of Soluble 4-Diphosphocytidyl-2-C-methyl-D-erythritol Kinase from Bacterial Pathogens

Cited by 23 publications

References 34 publications

Identification of novel small molecule inhibitors of 4-diphosphocytidyl-2-C-methyl-d-erythritol (CDP-ME) kinase of Gram-negative bacteria

Identification of novel small molecule inhibitors of 4-diphosphocytidyl-2-C-methyl-d-erythritol (CDP-ME) kinase of Gram-negative bacteria

Mechanistic Aspects of Carotenoid Biosynthesis

Formal synthesis of 4-diphosphocytidyl-2-C-methyl d-erythritol from d-(+)-arabitol

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