Mass spectral characterization of organophosphate-labeled lysine in peptides
Section snippets
Materials
Purified human serum albumin (essentially fatty acid free, cat. no. 05418), bovine actin (cat. no. 3653), human epidermal keratin (cat. no. K0253), DFP (cat. no. D0879), iodoacetamide (cat. no. I6125), Glu-fibrinopeptide B (cat. no. F3261), and mouse transferrin (cat. no. T0523) were obtained from Sigma–Aldrich–Fluka (St. Louis, MO, USA). Dithiothreitol (electrophoresis grade, cat. no. BP172-25) was obtained from Fisher Biotech (Fair Lawn, NJ, USA). Modified porcine trypsin (N-tosyl-l
OP-labeled peptides
A lysine from each of 22 peptides was found to have reacted with either CPO or DFP. Those 22 peptides came from seven different proteins: 5 peptides from human serum albumin, 2 peptides from human keratin 1, 1 peptide from human keratin 10, 6 peptides from bovine tubulin alpha, 1 peptide from bovine tubulin beta, 5 peptides from bovine actin, and 2 peptides from mouse transferrin (see Table 1).
When CPO reacts with lysine, the mass added to the peptide is 136 amu (for diethoxyphosphate). When DFP
Discussion
Reaction of OPs with tyrosine in a variety of proteins has been thoroughly documented [31]. We have now found a comparable reaction between OPs and lysine. Table 1 lists 22 peptides from seven proteins that are labeled on lysine. Involvement of this many different proteins strongly suggests that the reaction is commonplace. Nonetheless, we are aware of no other reports of OPs reacting with lysine on protein.
Analysis of the MS/MS spectra for 4 of the 22 peptides was presented. Comparable data
Acknowledgments
This work was supported by the U.S. Army Medical Research and Materiel Command (W81XWH-07-2-0034 to O.L.), the National Institutes of Health (U01 NS058056 to O.L. and P30CA36727 to Eppley Cancer Center), the Direction Générale de l’Armement of the French Ministry of Defense (DGA 03co010-05/PEA01 08 7 and DGA/PEA 08co501), and the Agence Nationale pour la Recherche (ANR-06-BLAN-0163). Mass spectra were obtained with the support of the Mass Spectrometry and Proteomics Core Facility at the
References (35)
- et al.
Clinical, biochemical, and neurobehavioural studies of workers engaged in the manufacture of quinalphos
Food Chem. Toxicol.
(2000) - et al.
Diethylphosphorylation of rat cardiac M2 muscarinic receptor by chlorpyrifos oxon in vitro
Toxicol. Lett.
(2001) - et al.
Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry assay for organophosphorus toxicants bound to human albumin at Tyr411
Anal. Biochem.
(2007) - et al.
Mass spectrometry identifies covalent binding of soman, sarin, chlorpyrifos oxon, diisopropyl fluorophosphate, and FP–biotin to tyrosines on tubulin: a potential mechanism of long term toxicity by organophosphorus agents
Chem. Biol. Interact.
(2008) - et al.
Reaction of a specific tyrosine residue of papain with diisopropylfluorophosphate
J. Biol. Chem.
(1969) - et al.
Covalent binding of the organophosphorus agent FP–biotin to tyrosine in eight proteins that have no active site serine
Chem. Biol. Interact.
(2009) - et al.
Chromosomal protein phosphorylation on basic amino acids
Methods Cell Biol.
(1978) Organophosphorus pesticides: do they all have the same mechanism of toxicity?
J. Toxicol. Environ. Health B
(1999)- et al.
Organophosphate toxicology: safety aspects of nonacetylcholinesterase secondary targets
Chem. Res. Toxicol.
(2004) - et al.
Acetylcholinesterase inhibition: does it explain the toxicity of organophosphorus compounds?
Arch. Toxicol.
(2006)
Review of health consequences from high-, intermediate-, and low-level exposure to organophosphorus nerve agents
J. Appl. Toxicol.
Organophosphorus ester-induced chronic neurotoxicity
Arch. Environ. Health
Neuropsychiatric evaluation in subjects chronically exposed to organophosphate pesticides
Toxicol. Sci.
Depression and pesticide exposures in female spouses of licensed pesticide applicators in the agricultural health study cohort
J. Occup. Environ. Med.
Neurologic symptoms in licensed pesticide applicators in the Agricultural Health Study
Hum. Exp. Toxicol.
Pesticide exposure and risk of Parkinson’s disease: a family-based case–control study
BMC Neurol.
Correlation of structure and function in enzyme action
Science
Cited by (44)
The cytoskeleton as a non-cholinergic target of organophosphate compounds
2021, Chemico-Biological InteractionsReal-time monitoring of fenitrothion in water samples using a silicon nanophotonic biosensor
2021, Analytica Chimica ActaPoisoning by organophosphorus nerve agents and pesticides: An overview of the principle strategies and current progress of mass spectrometry-based procedures for verification
2021, Journal of Mass Spectrometry and Advances in the Clinical LabCitation Excerpt :Reactive nucleophilic groups may include the hydroxyl-functions in the side chains of the amino acid serine, as relevant in AChE and butyrylcholinesterase (BChE) (Fig. 3), or tyrosine, as relevant in human serum albumin (HSA) (Section 2.4.3) [15]. In addition, amine-groups in lysine are also prone to phosphylation as shown for ubiquitin, albumin, keratin, tubulin, actin and transferrin [44–46]. OP-induced modifications of large proteins like BChE (molecular weight (MW), of monomer: 85 kDa) or HSA (MW of monomer 68 kDa) only cause slight changes of the MW (increase of about 0.115 kDa per phosphyl-group attached) or of the physicochemical properties (slightest alteration of the isoelectric point, pI, or hydrophobicity).
The cross-linking action of organophosphorus poisons; Implications for chronic neurotoxicity
2020, Handbook of Toxicology of Chemical Warfare Agents