Comparison of styrene-7,8-oxide adducts formed via reaction with cysteine, N-terminal valine and carboxylic acid residues in human, mouse and rat hemoglobin

Yeowell-O'Connell, Karen; Pauwels, W; Severi, Mario; Jin, Zuliang; Walker, Matthew R.; Rappaport, Stephen M.; Veulemans, Hendrik

doi:10.1016/s0009-2797(97)00059-8

Cited by 21 publications

(7 citation statements)

References 34 publications

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“…For these reasons the determination of SO adducts on cysteine using Raney nickel7,, 8 can possibly provide a better estimation of the Hb alkylation level. This finding may also explain previous reports14,, 15 indicating for SO a superior sensitivity in the determination of the cysteine adducts compared to those at the protein N‐terminus.…”

Section: Discussionsupporting

confidence: 85%

“…In this way the overall level of Hb alkylation is underestimated, as it has been shown that several different residues can be involved in the formation of SO adducts with Hb and albumin even at low levels of exposure 14,. 15 This finding also agrees with previous studies based on mass spectrometric analysis of the adducts SO‐Hb formed in vitro 16–18. However, a structural study of the interaction of SO with Hb at the level of the whole protein molecule has not yet been carried out, to our knowledge.…”

supporting

confidence: 91%

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Structural analysis of styrene oxide/haemoglobin adducts by mass spectrometry: identification of suitable biomarkers for human exposure evaluation

Basile

Ferranti

Mamone

et al. 2002

Rapid Comm Mass Spectrometry

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The structural characterisation of adducts formed by the in vitro reaction of haemoglobin (Hb) with styrene oxide (SO), the most reactive metabolite of the industrial reagent styrene, was obtained by liquid chromatography/electrospray ionisation mass spectrometry (LC/ES-MS) analysis of modified tryptic peptides of human Hb chains. The reactive sites of human Hb towards SO were identified through characterisation of alkylated tryptic peptides by matrix-assisted laser desorption/ionisation with tandem mass spectrometry (MALDI-MS/MS). A procedure was set up based on this characterisation, allowing Hb modification to be assessed by monitoring SO/Hb adducts using HPLC with selected ion recording (SIR) mass spectrometry. By this methodology it was also possible to compare advantages and disadvantages of presently available strategies for the measurement of Hb adducts with SO. The results obtained could most plausibly lead to the optimisation of molecular dosimetry of SO adducts, and the analytical procedure described herein could be applied to the biological monitoring of styrene exposure in the workplace.

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Section: Discussionsupporting

confidence: 85%

supporting

confidence: 91%

Structural analysis of styrene oxide/haemoglobin adducts by mass spectrometry: identification of suitable biomarkers for human exposure evaluation

Basile

Ferranti

Mamone

et al. 2002

Rapid Comm Mass Spectrometry

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“…An examination of the estimated second-order rate constants (Table ) reveals that BO is much more reactive with rat Hb than with any of the other nucleophiles examined and that the rank order of BO's reactivity toward Hb was rat > mouse > human. These findings are consistent with those observed for styrene 7,8-oxide ( , ) and ethylene oxide () and presumably reflect the presence of an additional reactive free cysteine residue in rat Hb (at position 125 on the β chain) which is not present in mice or humans (). Likewise, our results indicate that BO is about equally reactive with Alb from humans and rats, a finding which is consistent with the reactivity of 1,4-benzoquinone demonstrated by McDonald et al ().…”

Section: Discussionsupporting

confidence: 87%

“…This similar reactivity is probably related to the structural similarities of Alb molecules from both species, which contain a single free cysteine at position 34 ( , ). Overall, the reactivity of BO toward cysteinyl residues of Hb and Alb is comparable to that of styrene 7,8-oxide ( , ) and is substantially less than that of 1,4-benzoquinone ().…”

Section: Discussionmentioning

confidence: 96%

Formation of Hemoglobin and Albumin Adducts of Benzene Oxide in Mouse, Rat, and Human Blood

Lindstrom

Yeowell-O'Connell

Waidyanatha

et al. 1998

Chem. Res. Toxicol.

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Little is known about the formation and disposition of benzene oxide (BO), the initial metabolite arising from oxidation of benzene by cytochrome P450. In this study, reactions of BO with hemoglobin (Hb) and albumin (Alb) were investigated in blood from B6C3F1 mice, F344 rats, and humans in vitro. The estimated half-lives of BO in blood were 6.6 min (mice), 7.9 min (rats), and 7.2 min (humans). The following second-order rate constants were estimated for reactions between BO and cysteinyl residues of Hb and Alb [in units of L (g of Hb- or Alb-h)-1]: mouse Hb = 1.16 x 10(-)4, rat Hb = 15.4 x 10(-)4, human Hb = 0.177 x 10(-)4, mouse Alb = 2.68 x 10(-)4, rat Alb = 4.96 x 10(-)4, and human Alb = 5.19 x 10(-)4. These rate constants were used with BO-adduct measurements to assess the systemic doses of BO arising from benzene in vivo in published animal and human studies. Among rats receiving a single gavage dose of 400 mg of benzene/kg of body weight, the BO dose of 2.62 x 10(3) nM BO-h, predicted from Alb adducts, was quite similar to the reported AUC0-infinity = 1.09 x 10(3) nM BO-h of BO in blood. Interestingly, assays of Hb adducts in the same rats predicted a much higher dose of 14.7 x 10(3) nM BO-h, suggesting possible in situ generation of adducts within the erythrocyte. Doses of BO predicted from Alb adducts were similar in workers exposed to benzene [13.3 nM BO-h (mg of benzene/kg of body weight)-1] and in rats following a single gavage dose of benzene [8. 42 nM BO-h (mg of benzene/kg of body weight)-1]. Additional experiments indicated that crude isolates of Hb and Alb had significantly higher levels of BO adducts than dialyzed proteins, suggesting that conjugates of low-molecular-weight species were abundant in these isolates.

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“…4-6 It has been reported that SO can covalently bind to a variety of nucleophilic functional groups in proteins, including the sulfhydryl group of cysteine, imidazole of histidine, the carboxylic acid group of aspartic and glutamic acid, amino group of lysine, and the N -terminal amino group of proteins. 5,7-9 Among those nucleophilic amino acids, cysteine has been reported as the most reactive amino acid residue to be modified by SO. 8,10 Albumin and hemoglobin SO adducts have been identified both in humans and animals.…”

mentioning

confidence: 99%

Development of Enantioselective Polyclonal Antibodies to Detect Styrene Oxide Protein Adducts

et al. 2009

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Styrene has been reported to be pneumotoxic and hepatotoxic in humans and animals. Styrene oxide, a major reactive metabolite of styrene, has been found to form covalent binding with proteins, such as albumin and hemoglobin. Styrene oxide has two optical isomers and it was reported that the (R)-enantiomer was more toxic than the (S)-enantiomer. The purpose of this study was to develop polyclonal antibodies that can stereoselectively recognize proteins modified by styrene oxide enantiomers at cysteine residues. Immunogens were prepared by alkylation of thiolated keyhole limpet hemocyanin (KLH) with styrene oxide enantiomers. Polyclonal antibodies were raised by immunization of rabbits with the chiral immunogens. Titration tests showed all six rabbits generated high titers of antisera that recognize (R)- or (S)-coating antigens accordingly. No cross-reaction was observed toward the carrier protein (BSA). All three rabbits immunized with (R)-immunogen produced antibodies that show enantioselectivity to the corresponding antigen, while only one among the three rabbits immunized with (S)-immunogen generated antibodies with enantioselectivity of the recognition. The enantioselectivity was also observed in competitive ELISA and immunoblot analysis. Additionally, competitive ELISA tests showed that the immunorecognition required the hydroxyl group of the haptens. Immunoblot analysis demonstrated that the immunorecognition depended on the amount of protein adducts blotted and hapten loading in protein adducts. In summary, we successfully developed polyclonal antibodies to stereoselectively detect protein adducts modified by styrene oxide enantiomers.

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Comparison of styrene-7,8-oxide adducts formed via reaction with cysteine, N-terminal valine and carboxylic acid residues in human, mouse and rat hemoglobin

Cited by 21 publications

References 34 publications

Structural analysis of styrene oxide/haemoglobin adducts by mass spectrometry: identification of suitable biomarkers for human exposure evaluation

Structural analysis of styrene oxide/haemoglobin adducts by mass spectrometry: identification of suitable biomarkers for human exposure evaluation

Formation of Hemoglobin and Albumin Adducts of Benzene Oxide in Mouse, Rat, and Human Blood

Development of Enantioselective Polyclonal Antibodies to Detect Styrene Oxide Protein Adducts

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