Formation of DNA-protein cross-links in mammalian cells by levuglandin E2

Murthi, Krishna K.; Friedman, Libby R.; Oleinick, Nancy L.; Salomon, Robert G.

doi:10.1021/bi00066a034

Cited by 61 publications

(53 citation statements)

References 47 publications

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“…The mixture was applied to a C 18 SepPak cartridge (Waters) that had been preconditioned with methanol (5 ml) and water (10 ml). The SepPak was washed with heptane (10 ml) and heptane/ethyl acetate (1:1, v/v, 10 ml) before elution with methanol/ethyl acetate (2:3, v/v, 10 ml).…”

Section: Elisamentioning

confidence: 99%

Protein Adducts of Iso[4]levuglandin E2, a Product of the Isoprostane Pathway, in Oxidized Low Density Lipoprotein

Salomon

Wang

Brame

et al. 1999

Journal of Biological Chemistry

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Levuglandin (LG) E 2 , a cytotoxic seco prostanoic acid co-generated with prostaglandins by nonenzymatic rearrangements of the cyclooxygenase-derived endoperoxide, prostaglandin H 2 , avidly binds to proteins. That LGE 2 -protein adducts can also be generated nonenzymatically is demonstrated by their production during free radical-induced oxidation of low density lipoprotein (LDL). Like oxidized LDL, LGE 2 -LDL, but not native LDL, undergoes receptor-mediated uptake and impaired processing by macrophage cells. Since radicalinduced lipid oxidation produces isomers of prostaglandins, isoprostanes (isoPs), via endoperoxide intermediates, we postulated previously that a similar family of LG isomers, isoLGs, is cogenerated with isoPs. Now iso[4]LGE 2 -protein epitopes produced by radicalinduced oxidation of arachidonic acid in the presence of protein were detected with an enzyme-linked immunosorbent assay. Iso[4]LGE 2 -protein epitopes are also generated during free radical-induced oxidation of LDL. All of the LGE 2 isomers generated upon oxidation of LDL are efficiently sequestered by covalent adduction with LDL-based amino groups. The potent electrophilic reactivity of iso-LGs can be anticipated to have biological consequences beyond their obvious potential as markers for specific arachidonate-derived protein modifications that may be of value for the quantitative assessment of oxidative injury.

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Section: Elisamentioning

confidence: 99%

Protein Adducts of Iso[4]levuglandin E2, a Product of the Isoprostane Pathway, in Oxidized Low Density Lipoprotein

Salomon

Wang

Brame

et al. 1999

Journal of Biological Chemistry

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“…LGs, reactive byproducts of PG synthesis, have been reported to form covalent adducts with proteins ( 4-6 ), DNA ( 7,8 ), and phosphatidylethanolamine ( 9, 10 ). However, the oxidized forms of LGs, isoLGs, and isoprostanes have not been reported.…”

Section: Discussionmentioning

confidence: 99%

Identification of novel oxidized levuglandin D2 in marine red alga and mouse tissue

Kanai

Hiroki

Koshino

et al. 2011

Journal of Lipid Research

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“…Also, the initial species of the levuglandinyl adducts are known to be highly reactive. This can lead to cross-linking between proteins and DNA (4) and between proteins and small molecules that contain amine groups (6). In addition, levuglandinyl adducts can produce intermolecular cross-linking between proteins (3).…”

Section: Discussionmentioning

confidence: 99%

“…1). Levuglandins are known to react covalently with primary amines, such as the ⑀-amine of lysine, with proteins and with DNA (3,4). We have characterized the adducts that are formed by the reaction of lysine with LGE 2 or PGH 2 (2,5), and knowledge of their structures has provided a basis for analysis of the adducts in protein digests utilizing liquid chromatography-tandem mass spectrometry.…”

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confidence: 99%

Levuglandinyl Adducts of Proteins Are Formed via a Prostaglandin H2 Synthase-dependent Pathway after Platelet Activation

Boutaud

Zagol

et al. 2003

Journal of Biological Chemistry

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The product of oxygenation of arachidonic acid by the prostaglandin H synthases (PGHS), prostaglandin H 2 (PGH 2 ), undergoes rearrangement to the highly reactive ␥-ketoaldehydes, levuglandin (LG) E 2 , and LGD 2 . We have demonstrated previously that LGE 2 reacts with the ⑀-amine of lysine to form both the levuglandinyl-lysine Schiff base and the pyrrole-derived levuglandinyl-lysine lactam adducts. We also have reported that these levuglandinyl-lysine adducts are formed on purified PGHSs following the oxygenation of arachidonic acid. We now present evidence that the levuglandinyl-lysine lactam adduct is formed in human platelets upon activation with exogenous arachidonic acid or thrombin. After proteolytic digestion of the platelet proteins, and isolation of the adducted amino acid residues, this adduct was identified by liquid chromatography-tandem mass spectrometry. We also demonstrate that formation of these adducts is inhibited by indomethacin, a PGHS inhibitor, and is enhanced by an inhibitor of thromboxane synthase. These data establish that levuglandinyllysine adducts are formed via a PGHS-dependent pathway in whole cells, even in the presence of an enzyme that metabolizes PGH 2 . They also demonstrate that a physiological stimulus is sufficient to lead to the lipid modification of proteins through the levuglandin pathway in human platelets. Prostaglandin H synthase (PGHS)1 catalyzes the oxygenation of arachidonic acid to the endoperoxide, prostaglandin H 2 (PGH 2 ). PGH 2 is further metabolized to the prostanoids PGD 2 , PGE 2 , PGF 2a , thromboxane A 2 , and prostacyclin by specific enzymes. Also, PGH 2 in aqueous solutions undergoes non-enzymatic rearrangement to yield PGE 2 and PGD 2 , and 20% of it rearranges to the highly reactive ␥-ketoaldehydes, levuglandins (LG) E 2 and D 2 (1, 2) (Fig. 1). Levuglandins are known to react covalently with primary amines, such as the ⑀-amine of lysine, with proteins and with DNA (3, 4). We have characterized the adducts that are formed by the reaction of lysine with LGE 2 or PGH 2 (2, 5), and knowledge of their structures has provided a basis for analysis of the adducts in protein digests utilizing liquid chromatography-tandem mass spectrometry. Utilizing this analytical approach, we have demonstrated formation of LG-lysine adducts on PGHS-1 and PGHS-2 following the oxygenation of arachidonic acid (6). Formation of covalent adducts also was observed with proteins co-incubated with PGHS and arachidonic acid. These findings formed the basis for a hypothesis that PGHS activity in cells could generate levuglandinyl adducts of proteins.Oxygenation of arachidonic acid by PGHS-1 in platelet microsomes has been shown to produce arachidonic acid-derived adducts of multiple proteins (7). Such labeling also has been reported in whole platelets (8) and is increased when thromboxane A 2 synthase is inhibited. However, the reactive product of arachidonic acid that forms these protein adducts has not yet been characterized. We hypothesized that these adducts of platelet proteins...

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Formation of DNA-protein cross-links in mammalian cells by levuglandin E2

Cited by 61 publications

References 47 publications

Protein Adducts of Iso[4]levuglandin E2, a Product of the Isoprostane Pathway, in Oxidized Low Density Lipoprotein

Protein Adducts of Iso[4]levuglandin E2, a Product of the Isoprostane Pathway, in Oxidized Low Density Lipoprotein

Identification of novel oxidized levuglandin D2 in marine red alga and mouse tissue

Levuglandinyl Adducts of Proteins Are Formed via a Prostaglandin H2 Synthase-dependent Pathway after Platelet Activation

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