Lipoxin and Aspirin-Triggered 15-epi-Lipoxin Cellular Interactions Anti-Inflammatory Lipid Mediators

Serhan, Charles N.; Takano, Tomoko; Gronert, Karsten; Chiang, Nan; Clish, Clary B.

doi:10.1515/cclm.1999.052

Cited by 33 publications

(22 citation statements)

References 54 publications

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“…The chemical mechanism of the secondary step has been discussed for quite a while, but here we provide rigorous experimental evidence to prove its enzymatic nature: (i) pure 12S-LOXs (native and recombinant species) are capable of converting 12S-HpETE to HxA 3 ; (ii) heat denaturation abolished both their LOX and HxA 3 synthase activities; (iii) the reaction is highly stereoselective with respect to substrate (12R-HETE is no substrate) and product (8R-epimer is not formed); (iv) the basic kinetic parameters (Michaelis-Menten kinetics) are consistent with an enzymatic reaction; (v) 12S-LOX inhibitors, such as OPP, abolish hepoxilin A 3 formation from 12S-HpETE. As reported earlier, 5S-LOXs catalyze two consecutive steps during leukotriene biosynthesis (5-lipoxygenation of arachidonic acid to 5S-HpETE and secondary 5S-HpETE dehydration) (16), and multiple LOX reactions are involved in lipoxin formation (19). Here we report that 12S-LOXs can catalyze two consecutive reactions during hepoxilin biosynthesis.…”

Section: Discussionsupporting

confidence: 72%

“…Co-immunoprecipitation of 12S-LOX and HxA 3 Synthase Using an Anti-12S-LOX Antibody-Lipoxygenases are multifunctional enzymes that, in addition to their fatty acid oxygenase activity, exhibit a hydroperoxidase (14,15), a leukotriene synthase (16,17), and a lipoxin synthase activity (18,19). To test whether the leukocyte-type 12S-LOX of RINm5F cells may be involved in HxA 3 synthesis, we immunoprecipitated the enzyme from the cell lysis supernatant using an anti-12S-LOX antibody.…”

Section: Rat Insulinoma Rinm5f Cells Express a Leukocyte-type 12s-loxmentioning

confidence: 99%

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The Rat Leukocyte-type 12-Lipoxygenase Exhibits an Intrinsic Hepoxilin A3 Synthase Activity

Nigam

Patabhiraman

Ciccoli

et al. 2004

Journal of Biological Chemistry

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Hepoxilins are biologically relevant eicosanoids formed via the 12-lipoxygenase pathway of the arachidonic acid cascade. Although these eicosanoids exhibit a myriad of biological activities, their biosynthetic mechanism has not been investigated in detail. We examined the arachidonic acid metabolism of RINm5F rat insulinoma cells and found that they constitutively express a leukocyte-type 12S-lipoxygenase. Moreover, we observed that RINm5F cells exhibit an active hepoxilin A 3 synthase that converts exogenous 12S-HpETE (12S-5Z,8-Z,10E,14Z-12-hydro(pero)xy-eicosa-5,8,10,14-tetraenoic acid) or arachidonic acid predominantly to hepoxilin A 3 . 12S-lipoxygenase and hepoxilin A 3 synthase activities were co-localized in the cytosol; immunoprecipitation with an anti-12S-lipoxygenase antibody co-precipitated the two catalytic activities. These data suggested that hepoxilin A 3 synthase activity may be considered an intrinsic catalytic property of the leukocyte-type 12S-lipoxygenase. To test this hypothesis we cloned the leukocyte-type 12S-LOX from RINm5F cells, expressed it in Pichia pastoris, and found that the recombinant enzyme exhibited both 12S-lipoxygenase and hepoxilin A 3 synthase activities. The recombinant human platelet-type 12S-lipoxygenase and the porcine leukocyte-type 12S-lipoxygenase also exhibited hepoxilin A 3 synthase activity. In contrast, the native rabbit reticulocyte-type 15S-lipoxygenase did not convert 12S-HpETE to hepoxilin isomers. These data suggest that the positional specificity of lipoxygenases may be crucial for this catalytic function. This hypothesis was confirmed by site-directed mutagenesis studies that altered the positional specificity of the rat leukocyte-type 12S-and the rabbit reticulocyte-type 15-lipoxygenase. In summary, it may be concluded that naturally occurring 12S-lipoxygenases exhibit an intrinsic hepoxilin A 3 synthase activity that is minimal in lipoxygenase isoforms with different positional specificity.

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Section: Discussionsupporting

confidence: 72%

Section: Rat Insulinoma Rinm5f Cells Express a Leukocyte-type 12s-loxmentioning

confidence: 99%

The Rat Leukocyte-type 12-Lipoxygenase Exhibits an Intrinsic Hepoxilin A3 Synthase Activity

Nigam

Patabhiraman

Ciccoli

et al. 2004

Journal of Biological Chemistry

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“…LTA 4 is taken up by platelets and transformed to LXA 4 and LXB 4 via 12-LO activity (20). LXs act as anti-inflammatory and proresolution lipid mediators, by inhibiting both neutrophil and eosinophil transmigration into sites of infection and by promoting the noninflammatory infiltration of monocytes that is required for resolution and wound healing (21)(22)(23). LXs also stimulate macrophages to ingest and clear apoptotic neutrophils and elevate levels of the anti-inflammatory cytokine transforming growth factor beta 1 (24,25).…”

mentioning

confidence: 99%

5-Lipoxygenase Negatively Regulates Th1 Response during Brucella abortus Infection in Mice

et al. 2015

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Brucella abortus is a Gram-negative bacterium that survives inside host cells as a facultative intracellular pathogen (1). It infects humans and cattle, causing a chronic inflammatory disease known as brucellosis. In humans, brucellosis symptoms include undulant fever, endocarditis, arthritis, and osteomyelitis (2). In cattle, B. abortus causes miscarriage and infertility, leading to serious economic losses (3). The host immune response to B. abortus is initiated through innate immune mechanisms that recognize bacterial components and provide the necessary signals for the induction of an adaptive immune response (4). This specific adaptive immune response, which is Th1 mediated, plays a critical role in host control of B. abortus (5). The Th1-driven immune response to B. abortus involves CD4 ϩ and CD8 ϩ T lymphocytes, macrophages, dendritic cells (DCs), and proinflammatory cytokines such as gamma interferon (IFN-␥) and interleukin-12 (IL-12) (6-9).There is a growing interest in the immunoregulatory role of lipid mediators in infectious diseases (10-12). 5-Lipoxygenase (5-LO) is an enzyme required for the biosynthesis of two important groups of lipid mediators, leukotrienes (LTs) and lipoxins (LXs), both derived from arachidonic acid. 5-LO is made in several cell types, including neutrophils, eosinophils, monocytes/macrophages, dendritic cells, mast cells, and lymphocytes (13). To produce LTs, 5-LO acts on arachidonic acid, converting it to leukotriene A 4 (LTA 4 ). LTA 4 is then converted into leukotriene B 4 (LTB 4 ) or leukotriene C 4 (LTC 4 ), molecules that are exported from the cell (14). LTs are produced at the initial steps of the acute inflammatory response and act predominantly as proinflammatory lipid mediators. LTB 4 , for example, attracts neutrophils, monocytes, and lymphocytes to the site of inflammation and induces edema formation by increasing vascular permeability and plasma leakage at the site of inflammation (15-18). In contrast to LTs, production of LXs is dependent on cell-cell interaction by a process known as transcellular biosynthesis and requires the interaction between 5-LO and 15-or 12-lipoxygenase (15-LO or 12-LO, respectively) (19). Production of LXs by 5-15-LOs begins in eosinophils, monocytes, or epithelial cells (18). The 15-LO metabolic product, 15S-hydroperoxyeicosatetraenoic acid (15S-HPETE), is released by these cells, taken up by polymorphonuclear cells or monocytes, and then processed by 5-LO into bioactive lipoxin A 4 (LXA 4 ) or lipoxin B 4 (LXB 4 ). Production of LXs by 15-12-LO occurs after arachidonic acid conversion to LTA 4 by 5-LO in leukocytes. LTA 4 is taken up by platelets and transformed to LXA 4 and LXB 4 via 12-LO activity (20). LXs act as anti-inflammatory and proresolution lipid mediators, by inhibiting both neutrophil and eosinophil transmigration into sites of infection and by promoting the noninflammatory infiltration of monocytes that is required for resolution and wound healing (21-23). LXs also stimulate macrophages to ingest and clear apoptotic neutrop...

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“…The cysteinyl leukotrienes C 4 , D 4 , and E 4 increase vascular permeability and are effective constrictors of bronchial smooth muscle. 5-LO also participates in the formation of lipoxins, another group of arachidonate-derived bioactive lipids that are implicated in inflammatory and vascular events (2).…”

mentioning

confidence: 99%

The N-terminal Domain of 5-Lipoxygenase Binds Calcium and Mediates Calcium Stimulation of Enzyme Activity

Hammarberg¹,

Provost²,

Persson³

et al. 2000

Journal of Biological Chemistry

165

127

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The leukotrienes are important mediators in asthma as well as in other inflammatory and allergic disorders. 5-Lipoxygenase (5-LO 1 ; arachidonate:oxygen 5-oxidoreductase, EC 1.13.11.34) catalyzes two initial steps in the cellular production of leukotrienes. Thus, 5-LO converts arachidonic acid into 5(S)-hydroperoxy-6-trans-8,11,14-cis-eicosatetraenoic acid and subsequently into the unstable epoxide leukotriene A 4 , which, in turn, is the precursor of the biologically active leukotrienes B 4 , C 4 , D 4 , and E 4 (1). Leukotriene B 4 stimulates adherence of leukocytes to the vessel wall and is a potent chemotactic agent for these cells. The cysteinyl leukotrienes C 4 , D 4 , and E 4 increase vascular permeability and are effective constrictors of bronchial smooth muscle. 5-LO also participates in the formation of lipoxins, another group of arachidonate-derived bioactive lipids that are implicated in inflammatory and vascular events (2).Calcium is a well known 5-LO activator (for reviews, see Refs. 3-5). In brief, stimuli that elevate the intracellular Ca 2ϩ levels were shown to induce cellular 5-LO activity, and several reports have described Ca 2ϩ -induced translocation of 5-LO from the cytosol to cellular membranes. More detailed analyses showed an association primarily with the nuclear envelope, where the membrane-bound 5-LO-activating protein (FLAP) is also found and where the substrate arachidonic acid can be released from membrane lipids by cytosolic phospholipase A 2 (cPLA 2 ). The stimulatory effect of Ca 2ϩ is evident also for purified 5-LO. The basal enzyme activity, which is observed in the presence of a membrane fraction or lipids, increases up to 10-fold if micromolar concentrations of Ca 2ϩ are included in the assay mixture. 5-LO catalysis has been shown to occur at the lipid/water interface, and Ca 2ϩ -dependent binding of 5-LO to phospholipid vesicles has been reported. By several experimental approaches, we have recently demonstrated that 5-LO binds Ca 2ϩ in a reversible manner (3). A K d close to 6 M was determined by equilibrium dialysis, and the stoichiometry of maximum binding averaged around two Ca 2ϩ ions/5-LO molecule. We also showed that binding of calcium increased the hydrophobicity of 5-LO. Thus, a present conception is that calcium stimulates 5-LO activity and leukotriene production by promoting membrane association.The first structural determination of a mammalian 15-lipoxygenase (6) revealed that, similar to soybean lipoxygenases (7-9), it is composed of two major domains: a C-terminal domain containing the catalytic site and an N-terminal ␤-barrel domain. It seems reasonable that this is the overall structure also for 5-LO. The capability of 5-LO to bind more than one calcium ion and the calcium-dependent binding to phospholipids make 5-LO functionally similar to a group of calciumbinding proteins known as C2 domain proteins. The C2 domain is a conserved structural motif that forms an eight-stranded anti-parallel ␤-sandwich, and C2 domains have been identified in some 70 membrane-...

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Lipoxin and Aspirin-Triggered 15-epi-Lipoxin Cellular Interactions Anti-Inflammatory Lipid Mediators

Cited by 33 publications

References 54 publications

The Rat Leukocyte-type 12-Lipoxygenase Exhibits an Intrinsic Hepoxilin A3 Synthase Activity

The Rat Leukocyte-type 12-Lipoxygenase Exhibits an Intrinsic Hepoxilin A3 Synthase Activity

5-Lipoxygenase Negatively Regulates Th1 Response during Brucella abortus Infection in Mice

The N-terminal Domain of 5-Lipoxygenase Binds Calcium and Mediates Calcium Stimulation of Enzyme Activity

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