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

Abstract

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...

Increased levels of 4‐hydroxynonenal in human monocytes fed with malarial pigment hemozoin A possible clue for hemozoin toxicity

“…HNE is a strong electrophilic alkylating agent which spontaneously reacts with thiols, and less so with amino groups in amino acids, proteins and purine/pyrimidines [12][13][14][15]. HNE-mediated inhibition of numerous enzymes resides in blockage of functionally important thiol or amino groups [12][13][14][15].…”

“…A constant byproduct of lipoperoxidation is HNE, a strong electrophilic alkylating agent which spontaneously reacts with thiols and amino groups in proteins and purine/pyrimidines [12][13][14][15]. We report here a marked increase in membrane lipoperoxides and HNE levels in isolated hemozoin-fed human monocytes.…”

“…The parasite is unable to degrade heme, which aggregates to an insoluble polymer, called hemozoin [14]. Upon RBC rupture hemozoin is dispersed in the vasculature.…”

Biochemistry and pathology of radical-mediated protein oxidation

“…With some lipid carbonyls, Schiff-base formation may be of limited importance compared with Michael addition reactions [73,74]. In insulin (which lacks cysteine), histidine residues are selectively modified by Michael addition of 4-HNE [75]. With dialdehydes, Schiffbase formation and Michael addition appear to be simultaneous, thus cross-linking lysine residues [76].…”

Age-related alterations in oxidatively damaged proteins of mouse heart mitochondrial electron transport chain complexes

“…Protein modifications caused by ROS include the formation of lipid peroxidation adducts (4-hydroxynonenal or HNE and malondialdehyde or MDA), carbonylation of lysine, arginine, proline, and threonine, and nitration of tyrosine [28][29][30][31]. Oxidatively damaged proteins have been detected and identified by mass spectrometry [9,26,32].…”