Sulfur mustard alkylates steroid hormones and impacts hormone function in vitro

Lüling, Robin; Singer, Helena; Popp, Tanja; John, Harald; Boekhoff, Ingrid; Thiermann, Horst; Daumann, Lena J.; Karaghiosoff, Konstantin; Gudermann, Thomas; Steinritz, Dirk

doi:10.1007/s00204-019-02571-x

Cited by 8 publications

(4 citation statements)

References 39 publications

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“…Most recently, SM was used by the terrorist group islamic state of Iraq and the Levant (ISIL) in Syria and Northern Iraq, 3–6 and accidental exposures have also been reported, 7,8 thus underlining the need to understand the pathophysiology, to develop appropriate antidotes, and to establish analytical methods for biomedical verification of poisoning. Once SM enters the bloodstream, it forms adducts with various molecules by alkylation of, for example, proteins, DNA, hormones, glutathione, and N ‐acetylcysteine 9–14 . Human serum albumin (HSA) is a well‐known target of SM being alkylated at diverse amino acid residues including cysteine 34 (Cys 34 ) 3,8,14–16 .…”

Section: Introductionmentioning

confidence: 99%

Identification of creatine kinase and alpha‐1 antitrypsin as protein targets of alkylation by sulfur mustard

Lüling

Schmeißer²,

Siegert

et al. 2020

Drug Testing and Analysis

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Sulfur mustard (SM) is a toxic chemical warfare agent deployed in several conflicts within the last 100 years and still represents a threat in terroristic attacks and warfare. SM research focuses on understanding the pathophysiology of SM and identifying novel biomarkers of exposure. SM is known to alkylate nucleophilic moieties of endogenous proteins, for example, free thiol groups of cysteine residues. The two‐dimensional‐thiol‐differences in gel electrophoresis (2D‐thiol‐DIGE) technique is an initial proteomics approach to detect proteins with free cysteine residues. These amino acids are selectively labeled with infrared‐maleimide dyes visualized after GE. Cysteine residues derivatized by alkylating agents are no longer accessible for the maleimide–thiol coupling resulting in the loss of the fluorescent signal of the corresponding protein. To prove the applicability of 2D‐thiol‐DIGE, this technology was exemplarily applied to neat human serum albumin treated with SM, to lysates from human cell culture exposed to SM as well as to human plasma exposed to CEES (chloroethyl ethyl sulfide, an SM analogue). Exemplarily, the most prominent proteins modified by SM were identified by matrix‐assisted laser desorption/ionization time‐of‐flight (tandem) mass spectrometry, MALDI‐TOF MS(/MS), as creatine kinase (CK) from human cells and as alpha‐1 antitrypsin (A1AT) from plasma samples. Peptides containing the residue Cys282 of CK and Cys232 of A1AT were unambiguously identified by micro liquid chromatography‐electrospray ionization high‐resolution tandem‐mass spectrometry (μLC‐ESI MS/HR MS) as being alkylated by SM bearing the specific hydroxyethylthioethyl‐(HETE)‐moiety. Both peptides might represent potential biomarkers of SM exposure. This is the first report introducing these endogenous proteins as targets of SM alkylation.

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Section: Introductionmentioning

confidence: 99%

Identification of creatine kinase and alpha‐1 antitrypsin as protein targets of alkylation by sulfur mustard

Lüling

Schmeißer²,

Siegert

et al. 2020

Drug Testing and Analysis

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“…and Lilüng et. al [26]. previously presented the 1 H-NMR spectra of betamethasone, the active compound of Diphrophos, although their measurements were performed in d-DMSO and in a d-DMSO D2O mixture, hence a slight shift between their recorded spectra and the hereby measured one, which was performed in D2O, can be registered (Figure3/Diphrophos).…”

mentioning

confidence: 94%

Treatment of Hypertrophic Scars with Corticoid-Embedded Dissolving Microneedles

Decker

Szabó

Hoeksema

et al. 2022

Journal of Burn Care &Amp; Research

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Hypertrophic scarring (HTS) is frequently observed after deep dermal and full-thickness skin defects. Local drug delivery in HTS has been shown more effective compared to other (minimally) invasive treatments. Disadvantages being operator-dependency and non-uniform drug distribution. Moreover, injections are painful and difficult when confronted with extensive scars or HTS in children. Corticoid-embedded dissolving microneedles (CEDMN) were developed that provide painless skin penetration and direct dermal drug delivery. Hyaluronic acid-based DMN and CEDMN patches were utilized. Structural analysis was performed via nuclear magnetic resonance (NMR) spectroscopy while gel permeation chromatography (GPC) was applied to determine chain length (molar mass) and dispersity of hyaluronic acid. Mechanical properties were evaluated by compression testing. Five burn victims with HTS were included. For each individual, three comparable scars were chosen. One control scar was left untreated. Two scars were treated with either 600 or 800 µm CEDMN patches. Patients were treated monthly for 4 months. Treatment with 800 µm CEDMN was initiated after 8 weeks. Assessor-blinded POSAS was registered. Hydration, evaporation, color and elasticity were recorded. The physico-chemical characterization suggests that the mechanical properties enable skin penetration and adequate drug delivery. Patients experienced the therapy as painless. According to the POSAS, all scars improved over time. However, the scars that were treated with CEDMN patches improved faster and with increased increment. The 800 µm CEDMN ensured the fastest POSAS-decrease. Hyaluronic acid-based CEDMN patches are valuable alternatives to intracicatrical injections, as they offer a painless and effective method for administering corticosteroids in HTS.

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“…For verification of SM poisoning, protein‐adducts are well‐suited analytical target molecules because of their long half‐lives compared to those of hydrolysis and enzymatic biotransformation products 8,9 . As a highly electrophilic compound, SM reacts with nucleophilic groups of endogenous molecules like proteins, DNA bases, hormones, or glutathione, thus forming covalent modifications by addition of a characteristic hydroxyethylthioethyl (HETE)‐moiety 2,10–14 . In particular, free thiol‐groups of cysteines represent highly reactive targets for alkylation by SM.…”

Section: Introductionmentioning

confidence: 99%

“…8,9 As a highly electrophilic compound, SM reacts with nucleophilic groups of endogenous molecules like proteins, DNA bases, hormones, or glutathione, thus forming covalent modifications by addition of a characteristic hydroxyethylthioethyl (HETE)-moiety. 2,[10][11][12][13][14] In particular, free thiol-groups of cysteines represent highly reactive targets for alkylation by SM. Human serum albumin (HSA) contains a free cysteine residue at position 34 (Cys 34 ), which may be alkylated by SM.…”

Section: Introductionmentioning

confidence: 99%

Transthyretin as a target of alkylation and a potential biomarker for sulfur mustard poisoning: Electrophoretic and mass spectrometric identification and characterization

Schmeißer¹,

Lüling

Steinritz

et al. 2021

Drug Testing and Analysis

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For the verification of exposure to the banned blister agent sulfur mustard (SM) and the better understanding of its pathophysiology, protein adducts formed with endogenous proteins represent an important field of toxicological research. SM and its analogue 2-chloroethyl ethyl sulfide (CEES) are well known to alkylate nucleophilic amino acid side chains, for example, free-thiol groups of cysteine residues. The specific two-dimensional thiol difference gel electrophoresis (2D-thiol-DIGE) technique making use of maleimide dyes allows the staining of free cysteine residues in proteins. As a consequence of alkylation by, for example, SM or CEES, this staining intensity is reduced. 2D-thiol-DIGE analysis of human plasma incubated with CEES and subsequent matrix-assisted laser desorption/ionization time-of-flight (tandem) mass-spectrometry, MALDI-TOF MS(/MS), revealed transthyretin (TTR) as a target of alkylating agents. TTR was extracted from SM-treated plasma by immunomagnetic separation (IMS) and analyzed after tryptic cleavage by microbore liquid chromatography-electrospray ionization high-resolution tandem-mass spectrometry (μLC-ESI MS/HR MS). It was found that the Cys 10 -residue of TTR present in the hexapeptide C(-HETE)PLMVK was alkylated by the hydroxyethylthioethyl (HETE)moiety, which is characteristic for SM exposure. It was shown that alkylated TTR is stable in plasma in vitro at 37 C for at least 14 days. In addition, C(-HETE)PLMVK can be selectively detected, is stable in the autosampler over 24 h, and shows linearity in a broad concentration range from 15.63 μM to 2 mM SM in plasma in vitro. Accordingly, TTR might represent a complementary protein marker molecule for the verification of SM exposure.

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Sulfur mustard alkylates steroid hormones and impacts hormone function in vitro

Cited by 8 publications

References 39 publications

Identification of creatine kinase and alpha‐1 antitrypsin as protein targets of alkylation by sulfur mustard

Identification of creatine kinase and alpha‐1 antitrypsin as protein targets of alkylation by sulfur mustard

Treatment of Hypertrophic Scars with Corticoid-Embedded Dissolving Microneedles

Transthyretin as a target of alkylation and a potential biomarker for sulfur mustard poisoning: Electrophoretic and mass spectrometric identification and characterization

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