Determination of the Sites of 4-Hydroxy-2-nonenal Adduction to Protein by Electrospray Tandem Mass Spectrometry

Bolgar, Mark S.; Gaskell, Simon J.

doi:10.1021/ac9601021

Cited by 83 publications

(98 citation statements)

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“…Thus, His 5 was the modified residue when the addition of only one HNE molecule occurs, proven by the appearance of the b 5 ion containing one HNE moiety (Figure 3a), whereas both His 5 and His 10 were found to be modified in the case of di-alkylation (data not shown). As previously described by Bolgar and Gaskell [14], HNE-modified histidine residues give rise to an intense immonium ion at m/z 266 (noted H*), thus confirming the presence of an HNE molecule. Moreover, beside this typical immonium ion, a fragment ion corresponding to the protonated dehydrated HNE moiety can be clearly seen at m/z 139 in the product ion mass spectrum (Figure 3a).…”

Section: Hne-modified Oxidized Insulin B Chainsupporting

confidence: 80%

“…As described earlier, aldehydes are able to react with nitrogenous nucleophiles such as lysine, histidine, or cysteine to form Schiff bases (imines) whereas alkenals and hydroxy-alkenals could also undergo Michael (1-4) addition (Figure 1) [7][8][9][10][11]. Among these different lipid oxidation products, 4-hydroxy-2-nonenal (HNE) has received particular attention and was found to occur mainly on histidine or lysine residues via a Michael addition [8,[12][13][14]. Indeed, Bruenner et al have reported the mass spectrometric analysis of two HNE-modified model proteins (␤-lactoglobulin B and human hemoglobin) and thus showed, by comparison of the molecular masses, the preponderance of the Michael addition over the Schiff base condensation [7].…”

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confidence: 99%

“…Indeed, Bruenner et al have reported the mass spectrometric analysis of two HNE-modified model proteins (␤-lactoglobulin B and human hemoglobin) and thus showed, by comparison of the molecular masses, the preponderance of the Michael addition over the Schiff base condensation [7]. Moreover, the histidine residues were identified as the main target of HNE modification in human hemoglobin as well as ␤-lactoglobulin B [7], apomyoglobin [14], or subunit VIII of bovine heart cytochrome c oxidase [12]. In contrast, lysine residues seem to be the predominantly HNE-modified residues in glucose-6-phosphate dehydrogenase [15,16].…”

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confidence: 99%

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Study of protein modification by 4-hydroxy-2-nonenal and other short chain aldehydes analyzed by electrospray ionization tandem mass spectrometry

Fenaille

Guy

Tabet

2003

J. Am. Soc. Mass Spectrom.

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A convenient way to study lipid oxidation products-modified proteins by means of suitable model systems has been investigated. As a model peptide, the oxidized B chain of insulin has been chemically modified by either 4-hydroxy-2-nonenal (HNE) or hexanal and the extent, sites, and structure of modifications were assessed by electrospray mass spectrometry. A reduction step, using either NaCNBH 3 or NaBH 4 , was also studied to stabilize the alkylated compounds. From the data gathered, it appeared that NaCNBH 3 , when added at the beginning of incubation, dramatically influenced the HNE-induced modifications in terms of the addition mechanism (Schiff base formation instead of Michael addition) but also of the amino acid residues modified (N-terminal amino acid instead of histidine residues). However, by reducing the HNE-adducted species at the end of the reaction with NaBH 4 , the fragment ions obtained in the product ion scan experiments become more stable and thus, easier to interpret in terms of origin and mechanism involved. With regard to hexanal induced modifications, we have observed that hexanal addition under reductive conditions led to an extensive modification of the peptide backbone. Moreover, as confirmed by "in-source" collision followed by collision induced dissociation (CID) experiments on selected precursor ions (pseudo-MS 3 experiments), N,N-di-alkylations were first observed on the N-terminal residue and further on Lys 29 residue. On the other hand, compared to the native peptide, no significant changes in MS/MS fragmentation patterns (b and y ions series) were observed whatever the basic site modified by the aldehyde-addition. (J Am Soc Mass Spectrom 2003, 14, 215-226)

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Section: Hne-modified Oxidized Insulin B Chainsupporting

confidence: 80%

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confidence: 99%

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Study of protein modification by 4-hydroxy-2-nonenal and other short chain aldehydes analyzed by electrospray ionization tandem mass spectrometry

Fenaille

Guy

Tabet

2003

J. Am. Soc. Mass Spectrom.

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“…3. However, numerous attempts to detect Schiff base formation in various proteins have concluded that the extent to which HNE modifies Lys side chains is insignificant compared with the extent to which His side chains are modified (25,47,(61)(62)(63)(64)(65)(66). Moreover, immunohistochemical studies have shown that anti-HNE-His antibodies bind to amyloid plaques in AD brain tissue (26), but anti-HNE-Lys antibodies do not (36).…”

Section: Discussionmentioning

confidence: 99%

Membrane-mediated Amyloidogenesis and the Promotion of Oxidative Lipid Damage by Amyloid β Proteins

Murray

Liu

Komatsu

et al. 2007

Journal of Biological Chemistry

100

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Evidence of oxidative stress and the accumulation of fibrillar amyloid ␤ proteins (A␤) in senile plaques throughout the cerebral cortex are consistent features in the pathology of Alzheimer disease. To define a mechanistic link between these two processes, various aspects of the relationship between oxidative lipid membrane damage and amyloidogenesis were characterized by chemical and physical techniques. Earlier studies of this relationship demonstrated that oxidatively damaged synthetic lipid membranes promoted amyloidogenesis. The studies reported herein specify that 4-hydroxy-2-nonenal (HNE) is produced in both synthetic lipids and human brain lipid extracts by oxidative lipid damage and that it can account for accelerated amyloidogenesis. A␤ promotes the copper-mediated generation of HNE from polyunsaturated lipids, and in turn, HNE covalently modifies the histidine side chains of A␤. HNE-modified A␤ have an increased affinity for lipid membranes and an increased tendency to aggregate into amyloid fibrils. Thus, the prooxidant activity of A␤ leads to its own covalent modification and to accelerated amyloidogenesis. These results illustrate how lipid membranes may be involved in templating the pathological misfolding of A␤, and they suggest a possible chemical mechanism linking oxidative stress with amyloid formation. Alzheimer disease (AD)3 is an age-related neurodegenerative disorder characterized by misfolded and aggregated fibrillar amyloid ␤ proteins (A␤) in the brain. Among the factors associated with the pathogenesis of AD, oxidative stress is one of the most closely scrutinized (1, 2). It has been shown, for example, that the brain in AD has increased susceptibility to oxidative stress (3-5) and that isoprostanes, markers of oxidative stress, are specifically elevated (6 -8). Isoprostanes are chemically stable and nonreactive compounds that arise nonenzymatically from the spontaneous decomposition of lipid hydroperoxides. These hydroperoxides may decompose along other pathways, however, yielding highly reactive short chain alkenals such as 4-oxo-2-nonenal and 4-hydroxy-2-nonenal (HNE) (9 -17). HNE concentrations in human ventricular fluid are 8 -15 M and elevated in AD (3,18,19). HNE has a well known propensity to react with the side chains of various amino acid residues, and HNE-protein adducts have been used as biomarkers of oxidative stress (15).In light of these observations, it is noteworthy that lipid oxidation products such as HNE modify A␤ and increase A␤ misfolding (20 -25). Moreover, the immunoreactivity of antibodies to HNE-modified His residues localizes to amyloid plaques (26,27). This suggests that not only does A␤ promote lipid oxidation but that there may also be a mechanistic link between the lipid oxidation products formed during oxidative stress and A␤ misfolding (21). Conversely, several lines of evidence suggest that A␤ contribute to oxidative stress. For example, the overexpression of A␤ in transgenic mice, in Caenorhabditis elegans, and in cell culture results in an increase in...

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“…One important class of xenobiotic-protein adducts forms via lA-Michael addition of a ,,B-unsaturated carbonyls to nucleophilic amino acid residues. Many a,,B-unsaturated carbonyls including acrylic monomers, combustion by-products or metabolites such as acrolein [1], and endogenous compounds such as 4-hydroxynonenal react with protein nucleophilic side chains via Michael addition [9][10][11].Mass spectrometry has emerged as a powerful analytical tool for the characterization of post-translationally modified peptides and proteins because this approach can identify the modifying group and sites of modification [12]. Earlier studies demonstrated the utility of fast atom bombardment mass spectrometry (FAB/MS) for identification of specific thioether conjuAddress reprint requests to Dr. A. Daniel Jones, Facility for Adv anced Instrumen tation, University of California, Dav is, CA 95616.…”

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confidence: 99%

Fragmentation of protonated thioether conjugates of acrolein using low collision energies

Oberth

Jones

1997

J. Am. Soc. Mass Spectrom.

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The protonated mercapturic acid conjugate of acrolein, S-(3-oxopropyI)-N-acetyl-L-cysteine (I), undergoes facile retro-Michael loss of acrol ein in the gas phase. To determine whether extensive loss of acrolein would impede structural characterization of acrolein-peptide ad ducts, fragmentation reactions of a series of conjugates, formed by l,4-Michael addition of acrolein to peptides and cysteine derivatives, w ere investigated at collision cell potentials up to -50 V using a triple quadrupole mass spectrometer. Differences in fragmentation dynamics suggest protonation at the sulfur of the N-ac etylcysteine conjugate 1 facilitates retro-Michael elimination of acrolein with a low activation energy relative to other fragmentations. Analogous fragmentation was eliminated after borohydride reduction of the aldehyde to an alcohol. Retro-Michael fragmentation was not significant for acrolein conjugates of glutathione derivatives, suggesting that proton sequestration occurs in peptides with multiple amide linkages even when the peptide does not contain a basic amino group. An unexpected outcome of these experiments was the observation of a facile gas-phase cleavage of peptides on the N-terminal side of S-(3-oxopropylkysteine residues. Such fragmentation behavior may prove useful for locating cysteine residues in peptides. . Protein sulfhydryl groups are especially vulnerable to electrophiIic attack and form thioether conjugates whose presence indicates mechanisms of metabolic activation or detoxification. Knowledge of protein targets and sites of covalent attachment can be used to identify reactive metabolites, and these modified proteins can serve as biomarkers of exposure to xenobiotics in clinical and epidemiological studies [1][2][3][4][5][6][7][8]. One important class of xenobiotic-protein adducts forms via lA-Michael addition of a ,,B-unsaturated carbonyls to nucleophilic amino acid residues. Many a,,B-unsaturated carbonyls including acrylic monomers, combustion by-products or metabolites such as acrolein [1], and endogenous compounds such as 4-hydroxynonenal react with protein nucleophilic side chains via Michael addition [9][10][11].Mass spectrometry has emerged as a powerful analytical tool for the characterization of post-translationally modified peptides and proteins because this approach can identify the modifying group and sites of modification [12]. Earlier studies demonstrated the utility of fast atom bombardment mass spectrometry (FAB/MS) for identification of specific thioether conjuAddress reprint requests to Dr. A. Daniel Jones, Facility for Adv anced Instrumen tation, University of California, Dav is, CA 95616. E-mail: adjones@ucdavis.edu gates from biological samples [3,4,[13][14][15][16][17] . Using FAB or newer ionization techniques coupled with tandem mass spectrometry (MS/MS), researchers determined structures of thioether conjugates extracted from biological matrices [3][4][5][6][7][8][13][14][15][16][17][18][19][20][21][22]. In these MS/MS experiments, structural information was obtained fro...

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Determination of the Sites of 4-Hydroxy-2-nonenal Adduction to Protein by Electrospray Tandem Mass Spectrometry

Cited by 83 publications

References 26 publications

Study of protein modification by 4-hydroxy-2-nonenal and other short chain aldehydes analyzed by electrospray ionization tandem mass spectrometry

Study of protein modification by 4-hydroxy-2-nonenal and other short chain aldehydes analyzed by electrospray ionization tandem mass spectrometry

Membrane-mediated Amyloidogenesis and the Promotion of Oxidative Lipid Damage by Amyloid β Proteins

Fragmentation of protonated thioether conjugates of acrolein using low collision energies

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