Abstract
The hydrolysis of carbaryl by bovine serum albumin (BSA) was studied at toxicologically relevant concentrations (range 15–300 μM) in order to determine the role of this protein in the detoxication of the carbamate in vivo. The 1-naphthol released during the hydrolysis of carbaryl was monitored using gas chromatography coupled with mass spectrometry. BSA hydrolyzed carbaryl in a time-progressive way. The hydrolysis was also dependent of enzyme (1.0, 2.5, 5.0 and 7.0 mg ml−1) and substrate (range between 15 and 1,000 μM) concentration. The estimated turnover number and Michaelis–Menten constant were 1.6 × 10−4 s−1 and 430 μM, respectively. Thus, the second order rate constant was 0.37 M−1 s−1. At enzyme concentrations of 7.0 mg ml−1 and substrate concentrations ranging between 50 and 300 μM about 80% of substrate was hydrolyzed in 3 h. At lower substrate concentrations (15 and 30 μM carbaryl) also significant hydrolysis was detected at the highest enzyme concentration, even when these substrate concentrations were 30 and 15 times lower than the Michaelis–Menten constant. Although the efficacy of the enzymatic hydrolysis is low, the extrapolation of our results to the physiological albumin high concentrations (around 40 mg ml−1) suggests that the hydrolysis of carbaryl by serum albumins plays a critical role in the detoxication of this carbamate at in vivo toxicologically relevant concentrations.
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Abbreviations
- BSA:
-
Bovine serum albumin
- Carbaryl:
-
1-Naphthalenylmethyl carbamate
- Carbarylase:
-
Carbaryl hydrolyzing activity
- GC–MS:
-
Gas chromatography coupled with mass spectrometry
- HDCP:
-
O-Hexyl,O-2,5-dichlorophenyl phosphoramidate
- HSA:
-
Human serum albumin
- Paraoxon:
-
O,O′-Diethyl p-nitrophenyl phosphate
- RSA:
-
Rabbit serum albumin
References
Casida J, Augustinsson KB (1959a) Enzymic hydrolysis of N, N-dimethylcarbamoyl fluoride. Biochem Pharmacol 3:60–67
Casida J, Augustinsson KB (1959b) Reaction of plasma albumin with 1-naphthyl N-methylcarbamate and certain other esters. Biochim Biophys Acta 36:411–426
Chen-Goodspeed M, Sogorb MA, Wu F, Hong SB, Raushel FM (2001) Structural determinants of the substrate and stereochemical specificity of phosphotriesterase. Biochemistry 40(5):1325–1331
Doddamani HP, Ninnekar HZ (2001) Biodegradation of carbaryl by a Micrococcus species. Curr Microbiol 43(1):69–73
Hayatsu M, Nagata T (1993) Purification and characterization of carbaryl hydrolase from Blastobacter sp. strain M501. Appl Environ Microbiol 59(7):2121–2125
Karns JS, Tomasek PH (1991) Carbofuran hydrolases purification and properties. J Agric Food Chem 39(5):1004–1008
Li B, Sedlacek M, Manoharan I, Boopathy R, Duysen EG, Masson P, Lockridge O (2005) Butyrylcholinesterase, paraoxonase, and albumin esterase, but not carboxylesterase, are present in human plasma. Biochem Pharmacol 70:1673–1684
Maxwell DM, Lenz DE, Groff WA, Kaminskis A, Froehlich HL (1987) The effects of blood flow and detoxification on in vivo cholinesterase inhibition by soman in rats. Toxicol Appl Pharmacol 88:66–76
Means GE, Bender M (1975) Acetylation of human serum albumin by p-nitrophenyl acetate. Biochemistry 14:4989–4994
Mulbry WW, Eaton RW (1991) Purification and characterization of the N-methylcarbamate hydrolase from Pseudomonas strain CRL-OK. Appl Environ Microbiol 57(12):3679–3682
Peters T Jr (1996) Chemical aspects: albumin in medicine. In: Peters T (ed) All about albumin. Biochemistry, genetics, and medical applications. Academic Press Inc, San Diego pp 251–284
Pohlenz HD, Boidol W, Schuttke I, Streber WR (1992) Purification and properties of an Arthrobacter oxidans P52 carbamate hydrolase specific for the herbicide phenmedipham and nucleotide sequence of the corresponding gene. J Bacteriol 174(20):6600–6607
Smulders CJGM, Bueters TJH, Van Kleef RGDM, Vijverberg HPM (2003) Selective effects of carbamate pesticides on rat neuronal nicotinic acetylcholine receptors and rat brain acetylcholinesterase. Toxicol Appl Pharmacol 193:139–146
Sogorb MA, Vilanova E (2002) Enzymes involved in the detoxification of organophosphorus, carbamate and pyrethroid insecticides through hydrolysis. Toxicol Lett 128(1–3):215–228
Sogorb MA, Díaz-Alejo N, Escudero MA, Vilanova E (1998a) Phosphotriesterase activity identified in purified serum albumins. Arch Toxicol 72:219–226
Sogorb MA, Monroy A, Vilanova E (1998b) Chicken serum albumin hydrolyzes dichlorophenyl phosphoramidates by a mechanism based in a transient phosphorylation. Chem Res Toxicol 11(12):1441–1446
Sogorb MA, Sellero I, López-Rivadulla M, Céspedes V, Vilanova E (1999) EDTA-resistant and sensitive phosphotriesterase activities associated with albumin and lipoproteins in rabbit serum. Drug Metab Dispos 27(1):53–59
Sogorb MA, Carrera V, Benebent M, Vilanova E (2002) Rabbit serum albumin hydrolyzes the carbamate carbaryl. Chem Res Toxicol 15(4):520–526
Sogorb MA, Carrera V, Vilanova E (2004) Hydrolysis of carbaryl by human serum albumin. Arch Toxicol 78:629–634
Tildon JT, Ogilvie J (1972) The esterase activity of bovine mercaptalbumin. The reaction of the protein with p-nitrophenylacetate. J Biol Chem 247:1265–1271
World Health Organization (1986) Carbamates: a general introduction. In: World Health Organization (eds) Environmental health criteria 64. World Health Organization, Geneva
World Health Organization (1994) Carbaryl. In: World Health Organization (eds) Environmental health criteria 153. World Health Organization, Geneva
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The experiments performed for this work comply with the current laws of Spain.
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Sogorb, M.A., Álvarez-Escalante, C., Carrera, V. et al. An in vitro approach for demonstrating the critical role of serum albumin in the detoxication of the carbamate carbaryl at in vivo toxicologically relevant concentrations. Arch Toxicol 81, 113–119 (2007). https://doi.org/10.1007/s00204-006-0142-9
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DOI: https://doi.org/10.1007/s00204-006-0142-9