Abstract
A major challenge in organophosphate compound (OP) and OP nerve agent (OPNA) research has been in the identification and utilization of reliable biomarkers for rapid, sensitive, and efficient detection of OP exposure. Albumin has been widely studied as a biomarker for retrospective verification of exposure to OPNAs, including soman (GD), by detecting the phosphonylation of specific amino acid residues. The aim of the present study was to identify binding sites between GD and rabbit serum albumin in vitro and in vivo. A nano-liquid chromatography coupled with a quadrupole-orbitrap mass spectrometry (nLC-Q-Orbitrap-MS) was used to examine the GD-modified adducts of rabbit albumin. A total of 11 GD-modified sites were found in rabbit serum albumin across three experimental models. The following five GD-modified rabbit albumin sites, which were all lysine residues, were established in vivo: K188, K329, K162, K233, and K525. Two of these five lysine residues, K188 in peptide EK*ALISAAQER and K162 in peptide YK*AILTECCEAADK, were stable for at least 7 days in vivo. Molecular simulation of the GD–albumin interaction provided theoretical evidence for reactivity of the identified lysine residues. The findings suggest that these modifiable lysine residues are potential biomarkers of GD exposure for retrospective analysis by Q-Orbitrap-MS.
Similar content being viewed by others
References
Bajgar J (2004) Organophosphates/nerve agent poisoning: mechanism of action, diagnosis, prophylaxis, and treatment. Adv Clin Chem 38:151–216. https://doi.org/10.1016/s0065-2423(04)38006-6
Bao Y, Liu Q, Chen J, Lin Y, Wu B, Xie J (2012) Quantification of nerve agent adducts with albumin in rat plasma using liquid chromatography-isotope dilution tandem mass spectrometry. J Chromatogr A 1229:164–171. https://doi.org/10.1016/j.chroma.2012.01.032
Belinskaya DA, Shmurak VI, Prokofieva DS, Goncharov NV (2014) Serum albumin: search for new sites of interaction with organophosphorus compounds by the example of soman. Rus J Bioorg Chem 40(5):499–506. https://doi.org/10.1134/s1068162014050033
Black RM, Clarke RJ, Read RW, Reid MTJ (1994) Application of gas chromatography mass spectrometry and gas chromatography tandem mass spectrometry to the analysis of chemical warfare sample, found to contain residues of the nerve agent sarin sulfur mustard and their degradation products. J Chromatogr A 662(2):301–321. https://doi.org/10.1016/0021-9673(94)80518-0
Black RM, Harrison JM, Read RW (1999) The interaction of sarin and soman with plasma proteins: the identification of a novel phosphonylation site. Arch Toxicol 73(2):123–126. https://doi.org/10.1007/s002040050596
Carletti E, Colletier JP, Dupeux F, Trovaslet M, Masson P, Nachon F (2010) Structural evidence that human acetylcholinesterase inhibited by tabun ages through O-dealkylation. J Med Chem 53(10):4002–4008. https://doi.org/10.1021/jm901853b
Carter MD, Crow BS, Pantazides BG et al (2013) Direct quantitation of methyl phosphonate adducts to human serum butyrylcholinesterase by Immunomagnetic-UHPLC-MS/MS. Anal Chem 85(22):11106–11111. https://doi.org/10.1021/ac4029714
Chen S, Zhang J, Lumley L, Cashman JR (2013) Immunodetection of serum albumin adducts as biomarkers for organophosphorus exposure. J Pharmacol ExpTher 344(2):531–541. https://doi.org/10.1124/jpet.112.201368
Convention on the Prohibition of the Development, Production, Stockpiling and use of Chemical Weapons and their Destruction, Technical Secretariat of the Organisation for Prohibition of Chemical Weapons (1997) The Hague. Accessible through internet https://www.opcw.org/
Fidder A, Hulst AG, Noort D et al (2002) Retrospective detection of exposure to organophosphorus anti-cholinesterases: mass spectrometric analysis of phosphylated human butyrylcholinesterase. Chem Res Toxicol 15(4):582–590. https://doi.org/10.1021/tx0101806
Fu F, Sun F, Lu X et al (2018) A novel potential biomarker on Y263 site in human serum albumin poisoned by six nerve agents. J Chromatogr B Anal Technol Biomed Life Sci 1104:168–175. https://doi.org/10.1016/j.jchromb.2018.11.011
Gilley C, MacDonald M, Nachon F et al (2009) Nerve agent analogues that produce authentic soman, sarin, tabun, and cyclohexyl methylphosphonate modified human butyrylcholinesterase. Chem Res Toxicol 22(10):1680–1688. https://doi.org/10.1021/tx900090m
Jennings LL, Malecki M, Komives EA, Taylor P (2003) Direct analysis of the kinetic profiles of organophosphate-acetylcholinesterase adducts by MALDI-TOF mass spectrometry. Biochemistry 42(37):11083–11091. https://doi.org/10.1021/bi034756x
Jiang W, Dubrovskii YA, Podolskaya EP et al (2013) PHOS-select iron affinity beads enrich peptides for the detection of organophosphorus adducts on albumin. Chem Res Toxicol 26(12):1917–1925. https://doi.org/10.1021/tx400352h
John H, Breyer F, Thumfart JO, Hoechstetter H, Thiermann H (2010) Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) for detection and identification of albumin phosphylation by organophosphorus pesticides and G- and V-type nerve agents. Anal Bioanal Chem 398(6):2677–2691. https://doi.org/10.1007/s00216-010-4076-y
John H, Breyer F, Schmidt C, Mizaikoff B, Worek F, Thiermann H (2015) Small-scale purification of butyrylcholinesterase from human plasma and implementation of a LC-UV/ESI MS/MS method to detect its organophosphorus adducts. Drug Test Anal 7(10):947–956. https://doi.org/10.1002/dta.1792
Kranawetvogl A, Siegert M, Eyer F, Thiermann H, John H (2018) Verification of organophosphorus pesticide poisoning: detection of phosphorylated tyrosines and a cysteine-proline disulfide-adduct from human serum albumin after intoxication with dimethoate/omethoate. Toxicol Lett 299:11–20. https://doi.org/10.1016/j.toxlet.2018.08.013
Li B, Ricordel I, Schopfer LM et al (2010) Detection of adduct on tyrosine 411 of albumin in humans poisoned by Dichlorvos. Toxicol Sci 116(1):23–31. https://doi.org/10.1093/toxsci/kfq117
Majorek KA, Porebski PJ, Dayal A et al (2012) Structural and immunologic characterization of bovine, horse, and rabbit serum albumins. Mol Immunol 52(3–4):174–182
Marsillach J, Costa LG, Furlong CE (2013) Protein adducts as biomarkers of exposure to organophosphorus compounds. Toxicology 307:46–54. https://doi.org/10.1016/j.tox.2012.12.007
McDonough JH, Shih TM (1997) Neuropharmacological mechanisms of nerve agent-induced seizure and neuropathology. Neurosci Biobehav Rev 21(5):559–579. https://doi.org/10.1016/s0149-7634(96)00050-4
Mercey G, Verdelet T, Renou J et al (2012) Reactivators of acetylcholinesterase inhibited by organophosphorus nerve agents. Acc Chem Res 45(5):756–766. https://doi.org/10.1021/ar2002864
Molecular Operating Environment (MOE) (2018) 2018.01; Chemical Computing Group Inc., 1010 Sherbooke St. West, Suite #910, Montreal, QC, Canada, H3A 2R7
Ning YC (2018) Structural identification of organic compounds and organic spectroscopy. Science Press, Beijing
Noort D, Fidder A, van der Schans MJ, Hulst AG (2006) Verification of exposure to organophosphates: generic mass spectrometric method for detection of human butyrylcholinesterase adducts. Anal Chem 78(18):6640–6644. https://doi.org/10.1021/ac060954t
Pardio VT, Ibarra N, Rodriguez MA, Waliszewski KN (2001) Use of cholinesterase activity in monitoring organophosphate pesticide exposure of cattle produced in tropical areas. J Agri Food Chem 49(12):6057–6062. https://doi.org/10.1021/jf010431g
Peeples ES, Schopfer LM, Duysen EG et al (2005) Albumin, a new biomarker of organophosphorus toxicant exposure, identified by mass spectrometry. Toxicol Sci 83(2):303–312. https://doi.org/10.1093/toxsci/kfi023
Read RW, Riches JR, Stevens JA, Stubbs SJ, Black RM (2010) Biomarkers of organophosphorus nerve agent exposure: comparison of phosphylated butyrylcholinesterase and phosphylated albumin after oxime therapy. Arch Toxicol 84(1):25–36. https://doi.org/10.1007/s00204-009-0473-4
Schecter WP (2004) Cholinergic symptoms due to nerve agent attack: a strategy for management. Anesthesiol Clin N Am 22(3):579–590. https://doi.org/10.1016/j.atc.2004.04.005
Schmidt C, Breyer F, Blum M-M, Thiermann H, Worek F, John H (2014) V-type nerve agents phosphonylate ubiquitin at biologically relevant lysine residues and induce intramolecular cyclization by an isopeptide bond. Anal Bioanal Chem 406(21):5171–5185. https://doi.org/10.1007/s00216-014-7706-y
Shevchenko A, Tomas H, Havlis J, Olsen JV, Mann M (2006) In-gel digestion for mass spectrometric characterization of proteins and proteomes. Nat Protoc 1(6):2856–2860. https://doi.org/10.1038/nprot.2006.468
Silverstein RM, Webster FX, Kiemle DJ, Bryee DL (1991) Spectrometic identification of organic compounds. Wiley, New York
Sun F, Ding J, Yu H, Gao R, Wang H, Pei C (2016) Identification of new binding sites of human transferrin incubated with organophosphorus agents via Q Exactive LC-MS/MS. J Chromatogr B Anal Technol Biomed Life Sci 1022:256–264. https://doi.org/10.1016/j.jchromb.2016.04.028
Sun F, Ding J, Lu X et al (2017) Mass spectral characterization of tabun-labeled lysine biomarkers in albumin. J Chromatogr B Anal Technol Biomed Life Sci 1057:54–61. https://doi.org/10.1016/j.jchromb.2017.04.047
Tarhoni MH, Lister T, Ray DE, Carter WG (2008) Albumin binding as a potential biomarker of exposure to moderately low levels of organophosphorus pesticides. Biomarkers 13(4):343–363. https://doi.org/10.1080/13547500801973563
Thiede B, Lamer S, Mattow J et al (2000) Analysis of missed cleavage sites, tryptophan oxidation and N-terminal pyroglutamylation after in-gel tryptic digestion. Rapid Commun Mass Spectrom 14(6):496–502. https://doi.org/10.1002/(sici)1097-0231(20000331)14:6%3c496:Aid-rcm899%3e3.0.Co;2-1
Tsuchihashi H, Katagi M, Nishikawa M, Tatsuno M (1998) Identification of metabolites of nerve agent VX in serum collected from a victim. J Anal Toxicol 22(5):383–388. https://doi.org/10.1093/jat/22.5.383
Van der Schans MJ, Fidder A, van Oeveren D, Hulst AG, Noort D (2008) Verification of exposure to cholinesterase inhibitors: generic detection of OPCW schedule 1 nerve agent adducts to human butyrylcholinesterase. J Anal Toxicol 32(1):125–130
von der Wellen J, Winterhalter P, Siegert M, Eyer F, Thiermann H, John H (2018) A toolbox for microbore liquid chromatography tandem-high-resolution mass spectrometry analysis of albumin-adducts as novel biomarkers of organophosphorus pesticide poisoning. Toxicol Lett 292:46–54. https://doi.org/10.1016/j.toxlet.2018.04.025
Williams NH, Harrison JM, Read RW, Black RM (2007) Phosphylated tyrosine in albumin as a biomarker of exposure to organophosphorus nerve agents. Arch Toxicol 81(9):627–639. https://doi.org/10.1007/s00204-007-0191-8
Acknowledgements
This work was performed with the financial support of National Key R&D Program of China (Grant No. 2018YFC1602600).
Author information
Authors and Affiliations
Corresponding authors
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interests.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Fu, F., Gao, R., Zhang, R. et al. Verification of soman-related nerve agents via detection of phosphonylated adducts from rabbit albumin in vitro and in vivo. Arch Toxicol 93, 1853–1863 (2019). https://doi.org/10.1007/s00204-019-02485-8
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00204-019-02485-8