Using lysine adducts of human serum albumin to investigate the disposition of exogenous formaldehyde in human blood

Luca G. Regazzoni, Hasmik Grigoryan, Zhiying Ji, Xi Chen, Sarah I. Daniels, Deyin Huang, Sylvia Sanchez, Naijun Tang, Fenna Sille, Anthony T. Iavarone, Evan R. Williams, Luoping Zhang, Stephen M. Rappaport

Research output: Contribution to journalArticle

Abstract

Formaldehyde is a human carcinogen that readily binds to nucleophiles, including proteins and DNA. To investigate whether exogenous formaldehyde produces adducts in extracellular fluids, we characterized modifications to human serum albumin (HSA) following incubation of whole blood, plasma, and saliva with formaldehyde at concentrations of 1, 10 and 100 μM. The only HSA locus that showed the presence of formaldehyde modifications was Lys199. A N(6)-Lys adduct with added mass of 12 Da, representing a putative intramolecular crosslink, was detected in biological fluids that had been incubated with formaldehyde but not in control fluids. An adduct representing N(6)-Lys formylation was detected in all fluids, but levels did not increase above control values over the tested range of formaldehyde concentrations. An adduct representing N(6)-Lys199 acetylation was also measured in all samples. We then applied the assay to repeated samples of human plasma from 6 nonsmoking volunteer subjects (from Berkeley, CA), and single samples of serum from 15 workers exposed to airborne formaldehyde at about 1.5 ppm in a production facility and 15 control workers from Tianjin, China. Although all human plasma/serum samples contained basal levels of the products of N(6)-Lys formylation and acetylation, the putative crosslink product was not detected. Since the putative crosslink was observed in plasma incubated with formaldehyde at 1 μM, this suggests that the endogenous concentration of formaldehyde in serum was much lower than reported in the literature. Furthermore, concentrations of the formyl adduct were not higher in workers exposed to formaldehyde at about 1.5 ppm than in controls. Follow-up in vitro experiments with gaseous formaldehyde at 1.4 ppm detected the putative crosslink in plasma but not whole blood. This combination of results suggests that N(6) formylation occurs within cells with subsequent release of adducted HSA to the systemic circulation. Comparing across human samples, levels of N(6)-Lys199 formyl adducts were present at similar concentrations in subjects from California and China (about 1 mmol/mol HSA), but N(6)-Lys199 acetyl adducts were present at higher concentrations in Chinese subjects (0.34 vs. 0.13 mmol/mol HSA).

Original languageEnglish (US)
Pages (from-to)26-35
Number of pages10
JournalToxicology Letters
Volume268
DOIs
StatePublished - Feb 15 2017
Externally publishedYes

Fingerprint

Serum Albumin
Formaldehyde
Lysine
Blood
Plasma (human)
Acetylation
Fluids
Plasmas
China
Facility Regulation and Control
Serum
Nucleophiles
Extracellular Fluid
Saliva
Carcinogens
Volunteers
Assays
DNA

Keywords

  • Acetylation
  • Adducts
  • Blood
  • Covalent modification
  • Formaldehyde
  • Formylation
  • Lysine 199
  • Saliva

ASJC Scopus subject areas

  • Toxicology

Cite this

Regazzoni, L. G., Grigoryan, H., Ji, Z., Chen, X., Daniels, S. I., Huang, D., ... Rappaport, S. M. (2017). Using lysine adducts of human serum albumin to investigate the disposition of exogenous formaldehyde in human blood. Toxicology Letters, 268, 26-35. https://doi.org/10.1016/j.toxlet.2017.01.002

Using lysine adducts of human serum albumin to investigate the disposition of exogenous formaldehyde in human blood. / Regazzoni, Luca G.; Grigoryan, Hasmik; Ji, Zhiying; Chen, Xi; Daniels, Sarah I.; Huang, Deyin; Sanchez, Sylvia; Tang, Naijun; Sille, Fenna; Iavarone, Anthony T.; Williams, Evan R.; Zhang, Luoping; Rappaport, Stephen M.

In: Toxicology Letters, Vol. 268, 15.02.2017, p. 26-35.

Research output: Contribution to journalArticle

Regazzoni, LG, Grigoryan, H, Ji, Z, Chen, X, Daniels, SI, Huang, D, Sanchez, S, Tang, N, Sille, F, Iavarone, AT, Williams, ER, Zhang, L & Rappaport, SM 2017, 'Using lysine adducts of human serum albumin to investigate the disposition of exogenous formaldehyde in human blood', Toxicology Letters, vol. 268, pp. 26-35. https://doi.org/10.1016/j.toxlet.2017.01.002
Regazzoni, Luca G. ; Grigoryan, Hasmik ; Ji, Zhiying ; Chen, Xi ; Daniels, Sarah I. ; Huang, Deyin ; Sanchez, Sylvia ; Tang, Naijun ; Sille, Fenna ; Iavarone, Anthony T. ; Williams, Evan R. ; Zhang, Luoping ; Rappaport, Stephen M. / Using lysine adducts of human serum albumin to investigate the disposition of exogenous formaldehyde in human blood. In: Toxicology Letters. 2017 ; Vol. 268. pp. 26-35.
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abstract = "Formaldehyde is a human carcinogen that readily binds to nucleophiles, including proteins and DNA. To investigate whether exogenous formaldehyde produces adducts in extracellular fluids, we characterized modifications to human serum albumin (HSA) following incubation of whole blood, plasma, and saliva with formaldehyde at concentrations of 1, 10 and 100 μM. The only HSA locus that showed the presence of formaldehyde modifications was Lys199. A N(6)-Lys adduct with added mass of 12 Da, representing a putative intramolecular crosslink, was detected in biological fluids that had been incubated with formaldehyde but not in control fluids. An adduct representing N(6)-Lys formylation was detected in all fluids, but levels did not increase above control values over the tested range of formaldehyde concentrations. An adduct representing N(6)-Lys199 acetylation was also measured in all samples. We then applied the assay to repeated samples of human plasma from 6 nonsmoking volunteer subjects (from Berkeley, CA), and single samples of serum from 15 workers exposed to airborne formaldehyde at about 1.5 ppm in a production facility and 15 control workers from Tianjin, China. Although all human plasma/serum samples contained basal levels of the products of N(6)-Lys formylation and acetylation, the putative crosslink product was not detected. Since the putative crosslink was observed in plasma incubated with formaldehyde at 1 μM, this suggests that the endogenous concentration of formaldehyde in serum was much lower than reported in the literature. Furthermore, concentrations of the formyl adduct were not higher in workers exposed to formaldehyde at about 1.5 ppm than in controls. Follow-up in vitro experiments with gaseous formaldehyde at 1.4 ppm detected the putative crosslink in plasma but not whole blood. This combination of results suggests that N(6) formylation occurs within cells with subsequent release of adducted HSA to the systemic circulation. Comparing across human samples, levels of N(6)-Lys199 formyl adducts were present at similar concentrations in subjects from California and China (about 1 mmol/mol HSA), but N(6)-Lys199 acetyl adducts were present at higher concentrations in Chinese subjects (0.34 vs. 0.13 mmol/mol HSA).",
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AU - Daniels, Sarah I.

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AU - Sanchez, Sylvia

AU - Tang, Naijun

AU - Sille, Fenna

AU - Iavarone, Anthony T.

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N2 - Formaldehyde is a human carcinogen that readily binds to nucleophiles, including proteins and DNA. To investigate whether exogenous formaldehyde produces adducts in extracellular fluids, we characterized modifications to human serum albumin (HSA) following incubation of whole blood, plasma, and saliva with formaldehyde at concentrations of 1, 10 and 100 μM. The only HSA locus that showed the presence of formaldehyde modifications was Lys199. A N(6)-Lys adduct with added mass of 12 Da, representing a putative intramolecular crosslink, was detected in biological fluids that had been incubated with formaldehyde but not in control fluids. An adduct representing N(6)-Lys formylation was detected in all fluids, but levels did not increase above control values over the tested range of formaldehyde concentrations. An adduct representing N(6)-Lys199 acetylation was also measured in all samples. We then applied the assay to repeated samples of human plasma from 6 nonsmoking volunteer subjects (from Berkeley, CA), and single samples of serum from 15 workers exposed to airborne formaldehyde at about 1.5 ppm in a production facility and 15 control workers from Tianjin, China. Although all human plasma/serum samples contained basal levels of the products of N(6)-Lys formylation and acetylation, the putative crosslink product was not detected. Since the putative crosslink was observed in plasma incubated with formaldehyde at 1 μM, this suggests that the endogenous concentration of formaldehyde in serum was much lower than reported in the literature. Furthermore, concentrations of the formyl adduct were not higher in workers exposed to formaldehyde at about 1.5 ppm than in controls. Follow-up in vitro experiments with gaseous formaldehyde at 1.4 ppm detected the putative crosslink in plasma but not whole blood. This combination of results suggests that N(6) formylation occurs within cells with subsequent release of adducted HSA to the systemic circulation. Comparing across human samples, levels of N(6)-Lys199 formyl adducts were present at similar concentrations in subjects from California and China (about 1 mmol/mol HSA), but N(6)-Lys199 acetyl adducts were present at higher concentrations in Chinese subjects (0.34 vs. 0.13 mmol/mol HSA).

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