Detection of 4-hydroxynonenal as a product of lipid peroxidation in native ehrlich ascites tumor cells

Winkler, Paul; Lindner, Wolfgang; Esterbauer, Hermann; Schauenstein, E.; Schaur, R. J.; Khoschsorur, G.

doi:10.1016/0005-2760(84)90122-x

Cited by 51 publications

(15 citation statements)

References 9 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…(iii) The reaction of HNE with N-acetylhistidine is associated with the loss of a histidine residue and the stoichiometric formation of a product that, upon reduction with NaB[3H]H4, leads to incorporation of one equivalent of 3H into the product. Nevertheless, the Michael addition mechanism appears inconsistent with the fact that most of the N-acetylhistidine-HNE adduct does not react with 2,4-dinitrophenylhydrazine (2, to form a hydrazone (data not shown). Michael addition reactions should not affect the aldehyde function of HNE, unless the aldehyde moiety of the primary product reacts with the hydroxyl group at the C-4 position to form a hemiacetal (Fig.…”

Section: Methodsmentioning

confidence: 88%

“…The reaction of histidine residues with HNE provides the basis for methods by which the contributions of HNE in the modification of proteins can be determined. 4-Hydroxynonenal (HNE) is one of the major products of membrane peroxidation and has been shown to have a number of adverse biological effects such as high toxicity to mammalian cells (1,2), the lysis of erythrocytes (1), inactivation of enzymes (3), and inhibition ofthe synthesis of DNA and protein (3,4). These effects have been attributed in part to the fact that HNE reacts readily with sulfhydryl groups of proteins and low molecular weight metabolites, such as cysteine and glutathione, to yield stable thioether derivatives (5)(6)(7).…”

mentioning

confidence: 99%

See 1 more Smart Citation

Modification of histidine residues in proteins by reaction with 4-hydroxynonenal.

Uchida

Stadtman

1992

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

479

332

View full text Add to dashboard Cite

We find that histidine residues in proteins are major targets for reaction with the lipid peroxidation product 4-hydroxynon-2-enal (HNE). Reaction of insulin (which contains no sulfhydryl groups) with HNE leads to the generation of HNE-protein adducts, which are converted to radioactive derivatives upon subsequent treatment with NaB'H]1H4.Amino acid analysis of the modified protein showed that the HNE treatment leads to the selective loss of histdne residues and the stoichiometric formation of 3H-labeled amino acid derivatives. The same labeled products were detected in acid hydrolysates of polyhistidine and N-acetylhistidine after their reactions with HNE and NaB[3H]H4. The reaction of N-acetylhistidine with HNE led to the production of two compounds. Upon acid hydrolysis, both derivatives yielded stoichiometric amounts of histidine. However, after reduction with NaBH4, acid hydrolysis led to a mixture of amino acid derivatives [presumably, isomeric forms of Nff(NT)-1,4-dihydroxynonanylhistidine] that were indis hable from those obtained from insulin and polyhistidine after similar treatment. Although other possibilities are not excluded, it is suggested that the modification of hitidine residues in proteins by HNE involves a Michael-type addition ofthe imidazole nitrogen atom of histidine to the aj-unsaturated bond of HNE, followed by secondary reaction involving the aldehyde group with the C-4 hydroxyl group of HNE. The reaction of histidine residues with HNE provides the basis for methods by which the contributions of HNE in the modification of proteins can be determined.4-Hydroxynonenal (HNE) is one of the major products of membrane peroxidation and has been shown to have a number of adverse biological effects such as high toxicity to mammalian cells (1, 2), the lysis of erythrocytes (1), inactivation of enzymes (3), and inhibition ofthe synthesis of DNA and protein (3,4). These effects have been attributed in part to the fact that HNE reacts readily with sulfhydryl groups of proteins and low molecular weight metabolites, such as cysteine and glutathione, to yield stable thioether derivatives (5-7). Results of immunochemical studies with HNE-specific antibodies have shown that HNE-modified proteins are generated in vivo and, therefore, may play a physiological role (8-11). As noted by Esterbauer et al. (12), elucidation of the chemistry of the reactions of HNE with proteins is fundamental to an understanding of the mechanism of HNE cytotoxicity and also to the characterization of the epitopes in HNE-modified proteins that are recognized by antibodies.In the present paper, we demonstrate that, in addition to sulflhydryl groups and primary amino groups, the histidine residues of proteins are important targets for modification by HNE. In studies with insulin, which contains no sulfhydryl groups, it was shown that histidine is the only amino acid that is modified by exposure to HNE; the one lysine that is present in insulin was not modified under the same conditions. In addition, the results of model st...

show abstract

Section: Methodsmentioning

confidence: 88%

mentioning

confidence: 99%

Modification of histidine residues in proteins by reaction with 4-hydroxynonenal.

Uchida

Stadtman

1992

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

479

332

View full text Add to dashboard Cite

show abstract

“…HNE is an electrophilic lipid oxidation product and is one of the major constituents responsible for the toxic effects of lipid peroxidation (35). In addition to being cytotoxic, HNE promotes the activation of various signaling networks related to cell stress, including the DNA damage, endoplasmic reticulum stress, antioxidant, and heat shock response pathways (17).…”

Section: Discussionmentioning

confidence: 99%

Heat Shock Factor 1 Attenuates 4-Hydroxynonenal-mediated Apoptosis

Jacobs

Marnett

2007

Journal of Biological Chemistry

View full text Add to dashboard Cite

Lipid peroxidation is a consequence of both normal physiology and oxidative stress that generates various reactive metabolites, a principal end product being 4-hydroxynonenal (HNE). As a diffusible electrophile, HNE reacts extensively with cellular nucleophiles. Consequently, HNE alters cellular signaling and activates the intrinsic apoptotic cascade. We have previously demonstrated that in addition to promoting apoptosis, HNE activates stress response pathways, including the antioxidant, endoplasmic reticulum stress, DNA damage, and heat shock responses. Here we demonstrate that activation of the heat shock response by HNE is dependent on the expression and nuclear translocation of heat shock factor 1 (HSF1), which promotes the expression of heat shock protein 40 (Hsp40) and Hsp70-1. Ectopic expression and immunoprecipitation of c-Myc-tagged Hsp70-1 indicates that HNE disrupts the inhibitory interaction between Hsp70-1 and HSF1, leading to the activation heat shock gene expression. Using siRNA to silence HSF1 expression, we observe that HSF1 is necessary for the induction of Hsp40 and Hsp70-1 by HNE, and the lack of Hsp expression is correlated with an increase in apoptosis. Nrf2, the transcription factor that mediates the antioxidant response, was also silenced using siRNA. Silencing Nrf2 also enhanced the cytotoxicity of HNE, but not as effectively as HSF1. Silencing HSF1 expression facilitates the activation of JNK pro-apoptotic signaling and selectively decreases expression of the anti-apoptotic Bcl-2 family member Bcl-X L . Overexpression of Bcl-X L attenuates HNEmediated apoptosis in HSF1-silenced cells. Overall, activation of HSF1 and stabilization of Bcl-X L mediate a protective response that may contribute significantly to the cellular biology of lipid peroxidation.The oxidation of membrane phospholipids is an inescapable consequence of normal cellular respiration. A major route for lipid oxidation involves the abstraction of bis-allylic hydrogen atoms from polyunsaturated fatty acids by reactive oxygen species. The resulting lipid radicals react readily with dioxygen to form lipid hydroperoxides, which are mainly reduced to their corresponding hydroxyl species. A number of xenobiotic metabolites, anti-neoplastic drugs, transition metals, and disease states promote increased levels of both reactive oxygen species and oxidized lipids. The decomposition of oxidized lipids yields a variety of diffusible electrophiles, including the ␣,␤-unsaturated aldehyde, 4-hydroxynonenal (HNE).2 Diffusible lipid electrophiles are capable of modifying nucleophilic sites on DNA and proteins throughout the cell. Modification of cellular targets by HNE is found in multiple disease states, including atherosclerosis, ischemia-reperfusion injury, Parkinson disease, and Alzheimer disease (1-5). Plasma conjugates of HNE have been observed at low micromolar concentrations in healthy adults, implying the detectable generation of HNE under normal physiological conditions, although absolute steady-state concentrations are less clea...

show abstract

“…trans-4-Hydroxy-2-nonenal (HNE), a major product that is particularly damaging, exhibits toxicity to cells (6), lyses erythrocytes (7), and inhibits DNA and protein synthesis (8,9). HNE reacts with protein sulfhydryl groups to form thioethers (8,10), with primary amino groups, and with histidine imidazole groups (11).…”

mentioning

confidence: 99%

Inactivation of ethanol-inducible cytochrome P450 and other microsomal P450 isozymes by trans-4-hydroxy-2-nonenal, a major product of membrane lipid peroxidation.

Bestervelt

Vaz

Coon

1995

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

View full text Add to dashboard Cite

Of the microsomal P450 cytochromes, the ethanol-inducible isoform, P450 2E1, is believed to be predominant in leading to oxidative damage, including the generation of radical species that contribute to lipid peroxidation, and in the reductive j8-scission of lipid hydroperoxides to give hydrocarbons and aldehydes. In the present study, the sensitivity of a series of P450s to trans-4-hydroxy-2-nonenal (HNE), a known toxic product of membrane lipid peroxidation, was determined. After incubation of a purified cytochrome with HNE, the other components of the reconstituted system (NADPH-cytochrome P450 reductase, phosphatidylcholine, and NADPH) were added, and the rate of oxygenation of 1-phenylethanol to yield acetophenone was assayed. Inactivation occurs in a time-dependent and HNE concentrationdependent manner, with P450s 2E1 and lAl being the most sensitive, followed by isoforms 1A2, 3A6, and 2B4. At droperoxides to hydrocarbons and aldehydic acids-for example, the conversion of 13-hydroperoxy-9,11-octadecadienoic acid to pentane and 13-oxo-9,11-tridecadienoic acid-and the 2E1 cytochrome is one of the most active microsomal isoforms in this respect (4).The formation of reactive aldehydes, including alkenals, 2-alkenals, and 4-hydroxyalkenals, has been proposed to account for the cytotoxicity of lipid peroxidation products (5). trans-4-Hydroxy-2-nonenal (HNE), a major product that is particularly damaging, exhibits toxicity to cells (6), lyses erythrocytes (7), and inhibits DNA and protein synthesis (8, 9). HNE reacts with protein sulfhydryl groups to form thioethers (8, 10), with primary amino groups, and with histidine imidazole groups (11). Szweda et at (12) have described the inactivation of purified bacterial glucose-6-phosphate dehydrogenase by HNE accompanied by reaction with a lysine residue to give a stable secondary amine derivative, and more recently the formation of cross-linked protein less susceptible to proteolysis by the multicatalytic protease has been observed (13). In addition, Uchida and Stadtman have found that HNE inactivates purified rabbit muscle glyceraldehyde-3-phosphate dehydrogenase and at concentrations lower than 0.5 mM reacts primarily with cysteine and lysine residues (14).Since the first target of the toxic products of lipid peroxidation would be expected to be membranous enzymes, such as P450, we have undertaken a study of the sensitivity of hepatic microsomal P450 cytochromes to HNE. The present paper shows that the purified isozymes differ in the effect of HNE at various concentrations, with all of the cytochromes having decreased activity and with P450 2E1 being among the most strongly inactivated. Thus, the isoform that is particularly active in generating oxygen radicals that could initiate lipid peroxidation and in catalyzing the reductive cleavage of lipid hydroperoxides appears to be negatively regulated in the process. The observed inhibition of the various purified P450s and of P450 in microsomal membranes is of sufficient magnitude that it could cause significa...

show abstract

Detection of 4-hydroxynonenal as a product of lipid peroxidation in native ehrlich ascites tumor cells

Cited by 51 publications

References 9 publications

Modification of histidine residues in proteins by reaction with 4-hydroxynonenal.

Modification of histidine residues in proteins by reaction with 4-hydroxynonenal.

Heat Shock Factor 1 Attenuates 4-Hydroxynonenal-mediated Apoptosis

Inactivation of ethanol-inducible cytochrome P450 and other microsomal P450 isozymes by trans-4-hydroxy-2-nonenal, a major product of membrane lipid peroxidation.

Contact Info

Product

Resources

About