Phosphonylated tyrosine and lysine residues as biomarkers of local exposure of human hair to the organophosphorus nerve agents sarin and VX.
hair
keratin
mass spectrometry
protein adducts
verification
Journal
Drug testing and analysis
ISSN: 1942-7611
Titre abrégé: Drug Test Anal
Pays: England
ID NLM: 101483449
Informations de publication
Date de publication:
Jul 2023
Jul 2023
Historique:
revised:
19
01
2023
received:
21
11
2022
accepted:
10
02
2023
medline:
4
7
2023
pubmed:
15
2
2023
entrez:
14
2
2023
Statut:
ppublish
Résumé
We herein present for the first time a micro liquid chromatography-electrospray ionization high-resolution tandem-mass spectrometry (μLC-ESI MS/HR MS) procedure to detect phosphonylated tyrosine (Tyr) and lysine (Lys) residues obtained from human hair exposed to organophosphorus nerve agents (OPNA). In general, toxic OPNA react with endogenous blood proteins causing the formation of adducts representing well-known targets for biomedical analysis to prove exposure. In contrast, no protein-derived biomarker has been introduced so far to document local exposure of hair. Accordingly, we developed and characterized a μLC-ESI MS/HR MS method for the analysis of scalp hair exposed to OPNA in vitro. Type I and Type II keratin from hair was dissolved during lysis, precipitated and subjected to pronase-catalyzed hydrolysis yielding single adducted Lys and in a much higher amount Tyr residues. Exposure to sarin caused the adduction of an isopropyl methylphosphonic acid moiety and exposure to VX yielded adducts of ethyl methylphosphonic acid, well suited as biomarkers of exposure. These were of appropriate stability in the autosampler for 24 h. The biomarker yield obtained from hair of six individuals as well as from hair of six different parts of the body of one individual (armpit, beard, leg, arm, scalp, and pubic) differed reasonably indicating the variable individual protein composition and structure of hair. Exposed hair stored at ambient temperature for 9 weeks with contact to air and daylight showed stability of all adducts and therefore their suitability for verification of exposure.
Substances chimiques
Nerve Agents
0
Sarin
B4XG72QGFM
VX
9A4381183B
Lysine
K3Z4F929H6
Organophosphorus Compounds
0
Tyrosine
42HK56048U
Biomarkers
0
Chemical Warfare Agents
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
730-744Subventions
Organisme : Deutsche Forschungsgemeinschaft
ID : Grant/Award Number Research Training Group GRK 233
Organisme : German Research Foundation (Deutsche Forschungsgemeinschaft
ID : 2338
Informations de copyright
© 2023 The Authors. Drug Testing and Analysis published by John Wiley & Sons Ltd.
Références
John H, Balszuweit F, Steinritz D, Kehe K, Worek F, Thiermann H. Toxicokinetic aspects of nerve agents and vesicants. In: Gupta R, ed. Handbook of toxicology of chemical warfare agents. 3rd ed. San Diego: Elsevier Inc./Academic Press; 2020:875-929. doi:10.1016/B978-0-12-819090-6.00052-0
Chemical Weapons Convention: Convention on the prohibition of the development, production, stockpiling and use of chemical weapons and on their destruction. (2005) https://www.opcw.org/sites/default/files/documents/CWC/CWC_en.pdf. (accessed 15 October 2022). The Organisation for the Prohibition of Chemical Weapons (OPCW). The Hague. The Netherlands.
John H, van der Schans M, Koller M, et al. Fatal sarin poisoning in Syria 2013: forensic verification within an international laboratory network. Forensic Toxicol. 2018;36(1):61-71. doi:10.1007/s11419-017-0376-7
Statement by H.E. Ambassador Kenneth D. ward permanent representative of the United States of America to the OPCW in response to the director-general's presentation of the report of the OPCW investigation and identification team for Syria. https://www.opcw.org/sites/default/files/documents/2020/04/IIT%20Report%20USA%20National%20Statement%2004082020.pdf (accessed 15 October 2022). The Organisation for the Prohibition of Chemical Weapons (OPCW). The Hague. The Netherlands.
Tsuchihashi H, Katagi M, Nishikawa M. Identification of metabolites of nerve agent VX in serum collected from a victim. J Anal Toxicol. 1998;22(5):383-388. doi:10.1093/jat/22.5.383
Nakagawa T, Tu AT. Murders with VX: Aum Shinrikyo in Japan and the assassination of Kim Jong-Nam in Malaysia. Forensic Toxicol. 2018;36(2):542-544. doi:10.1007/s11419-018-0426-9
Vale JA, Marrs TC, Maynard RL. Novichok: a murderous nerve agent attack in the UK. Clin Toxicol. 2018;56(11):1093-1097. doi:10.1080/15563650.2018.1469759
Summary of the report on activities carried out in support of a request for technical assistance by the United Kingdom of Great Britain and Northern Ireland (Technical Assistance Visit TAV/02/18) S/1612/2018. https://www.opcw.org/sites/default/files/documents/S_series/2018/en/s-1612-2018_e___1_.pdf. (accessed 15 October 2022). The Organisation for the Prohibition of Chemical Weapons (OPCW). The Hague. The Netherlands.
Stone R. How German military scientists likely identified the nerve agent used to attack Alexei Navalny. Science https://www.sciencemag.org/news/2020/09/how-german-military-scientists-likely-identified-nerve-agent-used-attack-alexei-navalny, 10.1126/science.abe6561.
Summary of the report on activities carried out in support of a request for technical assistance by Germany (Technical Assistance Visit - TAV/01/20) S/1906/2020. https://www.opcw.org/sites/default/files/documents/2020/10/s-1906-2020%28e%29.pdf. (accessed 15 October 2022). The Organisation for the Prohibition of Chemical Weapons (OPCW). The Hague. The Netherlands.
Steindl D, Boehmerle W, Körner R, et al. Novichok nerve agent poisoning. Lancet. 2021;397(10270):249-252. doi:10.1016/S0140-6736(20)32644-1
Fidder A, Hulst AG, Noort D, et al. Retrospective detection of exposure to organophosphorus anti-cholinesterases: mass spectrometric analysis of phosphylated human butyrylcholinesterase. Chem Res Toxicol. 2002;15(4):582-590. doi:10.1021/tx0101806
John H, Breyer F, Schmidt C, Mizaikoff B, Worek F, Thiermann H. Small scale purification of butyrylcholinesterase from human plasma and implementation of a μLC-UV/ESI MS/MS method to detect its organophosphorus adducts. Drug Test Anal. 2015;7(10):947-956. doi:10.1002/dta.1792
John H, Breyer F, Thumfart JO, Höchstetter H, Thiermann H. 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. 2010;398(6):2677-2691. doi:10.1007/s00216-010-4076-y
Black R, Read RW. Biological markers of exposure to organophosphorus nerve agents. Arch Toxicol. 2013;87(3):421-437. doi:10.1007/s00204-012-1005-1
Peeples ES, Schopfer LM, Duysen EG, et al. Albumin, a new biomarker of organophosphorus toxicant exposure, identified by mass spectrometry. Toxicol Sci. 2005;83(2):303-312. doi:10.1093/toxsci/kfi023
von der Wellen J, Winterhalter P, Siegert M, Eyer F, Thiermann H, John H. A toolbox for liquid chromatography-tandem-mass spectrometry analysis of albumin-adducts as biomarkers of organophosphorus pesticide poisoning. Toxicol Lett. 2018;292:46-54. doi:10.1016/j.toxlet.2018.04.025
Kranawetvogl A, Worek F, Thiermann H, John H. Modification of human serum albumin by the nerve agent VX: microbore liquid chromatography/electrospray ionization high-resolution time-of-flight tandem mass spectrometry method for detection of phosphonylated tyrosine and novel cysteine containing disulfide adducts. Rapid Commun Mass Spectrom. 2016;30(19):2191-2200. doi:10.1002/rcm.7707
Kranawetvogl A, Küppers J, Siegert M, et al. Bioanalytical verification of V-type nerve agent exposure: simultaneous detection of phosphonylated tyrosines and cysteine-containing disulfide-adducts derived from human albumin. Anal Bioanal Chem. 2018;410(5):1463-1474. doi:10.1007/s00216-017-0787-7
Noort D, Hulst AG, van Zuylen A, van Rijssel E, van der Schans MJ. Covalent binding of organophosphorothioates to albumin: a new perspective for OP-pesticide biomonitoring? Arch Toxicol. 2009;83(11):1031-1036. doi:10.1007/s00204-009-0456-5
Li B, Eyer P, Eddleston M, Jiang W, Schopfer LM, Lockridge O. Protein tyrosine adduct in humans self-poisoned by chlorpyrifos. Toxicol Appl Pharmacol. 2013;269(3):215-225. doi:10.1016/j.taap.2013.03.021
Grigoryan H, Li B, Xue W, Grigoryan M, Schopfer LM, Lockridge O. Mass spectral characterization of organophosphate-labeled lysine in peptides. Anal Biochem. 2009;394(1):92-100. doi:10.1016/j.ab.2009.07.008
Schmidt C, Breyer F, Blum MM, Thiermann H, Worek F, John H. V-type nerve agents phosphonylate ubiquitin at biologically relevant lysine residues and induce intramolecular cyclization by an isopeptide bond. Anal Bioanal Chem. 2014;406(21):5171-5185. doi:10.1007/s00216-014-7706-y
Fu F, Guo F, Lu X, et al. Forensic analysis of soman exposure using characteristic fragment ions from protein adducts. Hum Exp Toxicol. 2021;40(9):1519-1527. doi:10.1177/09603271211001111
John H, Thiermann H. Poisoning by organophosphorus nerve agents and pesticides: an overview of the principle strategies and current progress of mass spectrometry-based procedures for verification. J Mass Spectrom Adv Clin Lab. 2021;19:20-31. doi:10.1016/j.jmsacl.2021.01.002
Schmeißer W, Siegert M, Thiermann H, Rein T, John H. Highly stable peptide adducts from hard keratins as biomarkers to verify local sulfur mustard exposure of hair by high-resolution mass spectrometry. Arch Toxicol. 2022;96(8):2287-2298. doi:10.1007/s00204-022-03307-0
International Air Transport Association (IATA). Dangerous goods regulation. 57thed. IATA, Montreal, Canada; 2016.
Schmeißer W, Lüling R, Steinritz D, Thiermann H, Rein T, John H. Transthyretin as a target of alkylation and a potential biomarker for sulfur mustard poisoning: electrophoretic and mass spectrometric identification and characterization. Drug Test Anal. 2022;14(1):80-91. doi:10.1002/dta.3146
Thermo Fisher Scientifc (accessed 18 October 2022) Instructions: In-Gel Tryptic Digestion Kit. https://tools.thermofisher.com/content/sfs/manuals/MAN0011497_InGel_Tryptic_Digest_UG.pdf
John H, Koller M, Worek F, Thiermann H, Siegert M. Forensic evidence of sulfur mustard exposure in real cases of human poisoning by detection of diverse albumin-derived protein-adducts. Arch Toxicol. 2019;93(7):1881-1891. doi:10.1007/s00204-019-02461-2
Siegert M, Gandor F, Kranawetvogl A, Börner H, Thiermann H, John H. Methionine329 in human serum albumin: a novel target for alkylation by sulfur mustard. Drug Test Anal. 2019;11(5):659-668. doi:10.1002/dta.2548
Blum MM, Richter A, Siegert M, Thiermann H, John H. Adducts of the blistering warfare agent sesquimustard with human serum albumin and their mass spectrometric identification for biomedical verification of exposure. Anal Bioanal Chem. 2020;412(28):7723-7737. doi:10.1007/s00216-020-02917-w
John H, Hörmann P, Schrader M, Thiermann H. Alkylated glutamic acid and histidine derived from protein-adducts indicate exposure to sulfur mustard in avian serum. Drug Test Anal. 2022;14(6):1140-1148. doi:10.1002/dta.3236
Zhang P, Chan W, Ang IL, et al. Revisiting fragmentation reactions of protonated α-amino acids by high-resolution electrospray ionization tandem mass spectrometry with collision-induced dissociation. Sci Rep. 2019;9(1):6453. doi:10.1038/s41598-019-42777-8
Kotiaho T, Eberlin MN, Vainiotalo P, Kostiainen R. Electrospray mass and tandem mass spectrometry identification of ozone oxidation products of amino acids and small peptides. J am Soc Mass Spectrom. 2000;11(6):526-535. doi:10.1016/S1044-0305(00)00116-1
Kranawetvogl A, Thiermann H, Siegert M, Eyer F, John H. 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. 2018;299:11-20. doi:10.1016/j.toxlet.2018.08.013
John H, Siegert M, Eyer F, Worek F, Thiermann H, Kranawetvogl A. Novel cysteine- and albumin-adduct biomarkers to prove human poisoning with the pesticide oxydemeton-S-methyl. Toxicol Lett. 2018;294:122-134. doi:10.1016/j.toxlet.2018.05.023
Gandor F, Gawlik M, Thiermann H, John H. Evidence of sulfur mustard exposure in human plasma by LC-ESI-MS-MS detection of the albumin-derived alkylated HETE-CP dipeptide and chromatographic investigation of its cis/trans isomerism. J Anal Toxicol. 2015;39(4):270-279. doi:10.1093/jat/bkv010
Fujii T, Takayama S, Ito Y. A novel purification procedure for keratin-associated proteins and keratin from human hair. J Biol Macromol. 2013;13(3):92-106. doi:10.14533/jbm.13.92
Robbins CR. Chemical and physical behavior of human hair. 5thed. Berlin Heidelberg: Springer; 2012. doi:10.1007/978-3-642-25611-0
Deb-Choudhury S, Plowman JE, Harland DP. Isolation and analysis of keratins and keratin-associated proteins from hair and wool. Methods Enzymol. 2016;568:279-301. doi:10.1016/bs.mie.2015.07.018
Langbein L, Rogers MA, Winter H, Praetzel S, Schweizer J. The catalog of human hair keratins. II. Expression of the six type II members in the hair follicle and the combined catalog of human type I and II keratins. J Biol Chem. 2001;276(37):35123-35132. doi:10.1074/jbc.M103305200
Schweizer J, Bowden PE, Coulombe PA, et al. New consensus nomenclature for mammalian keratins. J Cell Biol. 2006;174(2):169-174. doi:10.1083/jcb.200603161
Sweeny PJ, Walker JM. Pronase (EC 3.4.24.4). In: Burelli MM, ed. Methods in molecular biology. Vol.16. 1st ed. Totowa, NJ: Humana Press Inc; 1993:271-276. (Chapter 14)
Sigma-Aldrich. Product information sheet: Pronase. (accessed 16 October, 2022) https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/972/812/pronrobul.pdf
Lüling R, Schmeißer W, Siegert M, et al. Identification of creatine kinase and alpha-1 antitrypsin as protein targets of alkylation by sulfur mustard. Drug Test Anal. 2021;13(2):268-282. doi:10.1002/dta.2916
New York State department of health. Facts about nerve agents. (accessed 16 October, 2022) https://www.health.ny.gov/environmental/emergency/chemical_terrorism/docs/nerve_agents_tech.pdf
Leerunyakul K, Suchonwanit P. Asian hair: a review of structures, properties, and distinctive disorders. Clin Cosmet Investig Dermatol. 2020;13:309-318. doi:10.2147/CCID.S247390
Yang FC, Zhang Y, Rheinstädter MC. The structure of people's hair. PeerJ. 2:e619. doi:10.7717/peerj.619
Erdogan B. Anatomy and physiology of hair. In: Kutlubay Z, ed. Hair and Scalp Disorders. InTech; 2017:13-27. doi:10.5772/67269