Identification of an 8-Lipoxygenase Pathway in Nervous Tissue of Aplysia californica

Steel, Douglas; Tieman, Tamara L.; Schwartz, James H.; Feinmark, Steven J.

doi:10.1074/jbc.272.30.18673

Cited by 24 publications

(13 citation statements)

References 42 publications

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“…Lipids were extracted into ether and analyzed by reverse-phase HPLC as previously described (65). The 12-HETE was quantified by UV absorbance at 237 nm.…”

Section: Figurementioning

confidence: 99%

Complement-independent Ab-induced peroxide lysis of platelets requires 12-lipoxygenase and a platelet NADPH oxidase pathway

Nardi

Feinmark²,

Hu³

et al. 2004

J. Clin. Invest.

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“…Lipids were extracted into ether and analyzed by reverse-phase HPLC as previously described (65). The 12-HETE was quantified by UV absorbance at 237 nm.…”

Section: Figurementioning

confidence: 99%

Complement-independent Ab-induced peroxide lysis of platelets requires 12-lipoxygenase and a platelet NADPH oxidase pathway

Nardi

Feinmark²,

Hu³

et al. 2004

J. Clin. Invest.

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“…These enzymes oxygenate arachidonate in the 5R, 8R, 11R, or 12R configurations. The arachidonate (8R)-lipoxygenases have been studied in detail from two corals, Pseudoplexaura porosa and P. homomalla (46 -48), and from nervous tissue of Aplysia californica (43). Brash and co-workers (3) cloned and sequenced the (8R)-lipoxygenase from P. homomalla and found that it belonged to the lipoxygenase gene family.…”

Section: Different Strains Of Take-allmentioning

confidence: 99%

“…c Based on formation of 13-H(P)OTrE(n-3) only. (3,43,44). These enzymes oxygenate arachidonate in the 5R, 8R, 11R, or 12R configurations.…”

Section: Different Strains Of Take-allmentioning

confidence: 99%

Manganese Lipoxygenase

Oliw

1998

Journal of Biological Chemistry

165

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A linoleic acid (13R)-lipoxygenase was purified to homogeneity from the culture medium of Gä umannomyces graminis, the take-all fungus, by hydrophobic interaction, cation exchange, lectin affinity, and size-exclusion chromatography. The purified dioxygenase lacked light absorption between 300 and 700 nm. Gel filtration indicated an apparent molecular mass of ϳ135 kDa in 6 M urea and ϳ160 kDa in buffer. SDS-polyacrylamide gel electrophoresis (PAGE) showed that the enzyme was heterogeneous in size and consisted of diffuse protein bands of 100 -140 kDa. Treatment with glycosidases for N-and O-linked oligosaccharides yielded a distinct protein of ϳ73 kDa on SDS-PAGE. Atomic emission spectroscopy indicated 0.5-1.0 manganese atom/enzyme molecule. The isoelectric point was ϳ9.7, and the enzyme was active between pH 5 and 11 with optimum activity at pH 7.0. For molecular oxygen, K m was 30 M and V max 10 mol mg . The enzyme oxidized linolenic acid twice as fast as linoleic acid. The main products were identified by mass spectrometry as 13-hydroperoxy-(9Z,11E,15Z)-octadecatrienoic and 13-hydroperoxy-(9Z,11E)-octadecadienoic acids, respectively. After reduction of the hydroperoxide, steric analysis of methyl 13-hydroxyoctadecadienoate by chiral high performance liquid chromatography yielded one enantiomer (>95%), which co-eluted with the R-stereoisomer of methyl (13R,13S)-hydroxyoctadecadienoate. Arachidonic and dihomogammalinolenic acids were not substrates, while oxygen consumption, UV analysis, and mass spectrometric analysis indicated that ␥-linolenic acid was oxygenated both at C-11 and C-13. The enzyme was active at 60°C and after treatment with 6 M urea. It was strongly inhibited by 10 -50 M concentrations of eicosatetraynoic acid and a lipoxygenase inhibitor (N-(3-phenoxycinnamyl)acetohydroxamic acid), but many other lipoxygenase inhibitors (100 M) were without effect. We conclude that, after deglycosylation, the enzyme has the same size on SDS-PAGE as mammalian and marine lipoxygenases, but it differs from all previously described lipoxygenases in three ways. It is secreted, it forms (13R)-hydroperoxy-(9Z,11E)-octadecadienoic acid, and it contains manganese.

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“…Prior to analysis, samples were dried under a stream of nitrogen and reconstituted in ethyl acetate. Gas chromatography-mass spectrometry (GC-MS) analysis was performed as described previously (53), using a Hewlett Packard 5987A GC-MS instrument equipped with a DB-1 fused-silica capillary column (30 m by 0.2 mm), using helium as a carrier gas at injector and source temperatures of 220°C and 200°C, respectively. Samples were ionized by electron impact (70 eV).…”

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confidence: 99%

Gemfibrozil Inhibits Legionella pneumophila and Mycobacterium tuberculosis Enoyl Coenzyme A Reductases and Blocks Intracellular Growth of These Bacteria in Macrophages

Reich-Slotky¹,

Kabbash

Della‐Latta³

et al. 2009

J Bacteriol

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We report here that gemfibrozil (GFZ) inhibits axenic and intracellular growth of Legionella pneumophila and of 27 strains of wild-type and multidrug-resistant Mycobacterium tuberculosis in bacteriological medium and in human and mouse macrophages, respectively. At a concentration of 0.4 mM, GFZ completely inhibited L. pneumophila fatty acid synthesis, while at 0.12 mM it promoted cytoplasmic accumulation of polyhydroxybutyrate. To assess the mechanism(s) of these effects, we cloned an L. pneumophila FabI enoyl reductase homolog that complemented for growth an Escherichia coli strain carrying a temperature-sensitive enoyl reductase and rendered the complemented E. coli strain sensitive to GFZ at the nonpermissive temperature. GFZ noncompetitively inhibited this L. pneumophila FabI homolog, as well as M. tuberculosis InhA and E. coli FabI.The advent of AIDS and the emergence of many multidrugresistant bacterial species have led to renewed efforts to find new antibiotics. The most commonly used antibiotics act by blocking bacterium-specific DNA, RNA, or protein synthesis. Mycobacterium tuberculosis is a major exception to this generalization. While streptomycin, an inhibitor of bacterial protein synthesis, was the first antibiotic to be used successfully to treat M. tuberculosis, isoniazid (INH), an inhibitor of mycobacterial lipid synthesis, is presently the drug most commonly used to treat infections with this organism (2, 43). The differential sensitivity to INH of M. tuberculosis versus mammalian cells reflects the fact that most bacterial fatty acid synthases (type II synthases) are comprised of discrete, separable enzymes encoded by separate genes, while mammalian fatty acid synthases (type I) are dimeric proteins in which a single polypeptide catalyzes the seven enzymatic activities of fatty acid synthesis (21, 52).In previous studies (45), we reported that gemfibrozil (GFZ), a commonly prescribed and well-tolerated hypolipidemic drug, inhibits the export of various organic anions, including penicillin and fluoroquinolones, from murine macrophages, thereby elevating the intracellular concentration of these antibiotics and enhancing their capacity to block intracellular growth of Listeria monocytogenes. In exploring this system, we discovered that while GFZ has no effect on axenic or intracellular growth of Listeria monocytogenes, it inhibits axenic growth of all Legionella pneumophila strains tested and of 5 wild-type and 22 multidrug-resistant strains of M. tuberculosis and inhibits intracellular growth of L. pneumophila Philadelphia-1 and M. tuberculosis H37RV in human and mouse macrophages, respectively.Both M. tuberculosis and L. pneumophila are facultative intracellular pathogens that enter host macrophages by phagocytosis (25, 26), grow in nonlysosomal membrane-bound cytoplasmic vacuoles (24), have special nutrient requirements (38, 54, 55), and produce a relatively unique spectrum of membrane lipids (7, 57). However, M. tuberculosis is a slow-growing and dangerous organism with which to work. In cont...

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Identification of an 8-Lipoxygenase Pathway in Nervous Tissue of Aplysia californica

Cited by 24 publications

References 42 publications

Complement-independent Ab-induced peroxide lysis of platelets requires 12-lipoxygenase and a platelet NADPH oxidase pathway

Complement-independent Ab-induced peroxide lysis of platelets requires 12-lipoxygenase and a platelet NADPH oxidase pathway

Manganese Lipoxygenase

Gemfibrozil Inhibits Legionella pneumophila and Mycobacterium tuberculosis Enoyl Coenzyme A Reductases and Blocks Intracellular Growth of These Bacteria in Macrophages

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