Development of an automated on-line pepsin digestion–liquid chromatography–tandem mass spectrometry configuration for the rapid analysis of protein adducts of chemical warfare agents

Carol-Visser, Jeroen; Schans, Marcel van der; Fidder, A.; Hulst, A.G.; Baar, Ben L. M. van; Irth, H.; Noort, Daan

doi:10.1016/j.jchromb.2008.06.008

Cited by 45 publications

(27 citation statements)

References 23 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Besides free cysteine residues, the highly reactive electrophile SM is known to react with other amino acid residues containing nucleophilic moieties such as Glu, 12 His, 11 as well as the N-terminus in proteins. Accordingly, exploring novel biomarkers for SM intoxication is still an important field of research.…”

Section: Resultsmentioning

confidence: 99%

“…2,[11][12][13][14] By alkylation and subsequent hydrolysis, the characteristic hydroxyethylthioethyl (HETE) moiety is attached. 2,[11][12][13][14] By alkylation and subsequent hydrolysis, the characteristic hydroxyethylthioethyl (HETE) moiety is attached.…”

mentioning

confidence: 99%

“…Therefore, toxicity of SM is assumed to be due to the alkylation of proteins and DNA resulting in the functional impairment of cells, tissues, organs, and physiological processes. 12,[17][18][19][20][21][22][23] Based on the proteolysis of alkylated HSA, several analytical methods targeting the Cys 34 modification as HETE-CysProPhe (HETE-CPF) and HETE-CysPro (HETE-CP) were developed and have proven to be reliable biomarkers of exposure in vitro, in vivo, and in real cases of SM poisoning. 2 For the verification of an alleged use of SM, protein adducts reveal beneficial long-term stability of several weeks or even months in vivo when compared to products of SM hydrolysis and enzymatic biotransformation.…”

mentioning

confidence: 99%

“…Due to its bifunctional structure, crosslinks between biomolecules may occur. 12,[15][16][17][18][19][20][21][22][23][24] Herein, we present Met 329 as an additional target of SM in HSA. 15,16 Most relevant biomarkers are alkylated peptides derived from human serum albumin (HSA).…”

mentioning

confidence: 99%

See 3 more Smart Citations

Methionine³²⁹ in human serum albumin: A novel target for alkylation by sulfur mustard

Siegert

Gandor

Kranawetvogl

et al. 2019

Drug Testing and Analysis

View full text Add to dashboard Cite

Exposure to the vesicant sulfur mustard (SM) may lead to erythema and blistering.Toxicity of SM is hypothesized due to the alkylation of DNA bases and nucleophilic amino acid side chains in proteins (adducts) by forming the hydroxyethylthioethyl (HETE) moiety. Despite its prohibition by the chemical weapons convention, SM still represents a serious threat to military personnel and civilians. Therefore, development and improvement of forensic analytical methods for the verification of SM exposure is of high interest. Protein adducts have been shown to be highly suitable and beneficial biomarkers of poisoning. Herein we present methionine 329 in human serum albumin (HSA) as a novel target of SM forming a HETE-methionyl sulfonium ion.The alkylated tetrapeptide LeuGlyMet 329 (-HETE)Phe, LGM(-HETE)F, was detected after pepsin-mediated proteolysis and subsequent analysis by microbore liquid chromatography-electrospray ionization-high-resolution tandem-mass spectrometry.Compound identity was confirmed by a synthetic reference. Proteolysis conditions for HSA were optimized towards maximum yield of LGM(-HETE)F and its limit of identification (32.3 nM SM in serum) was similar to those of the established HSA-derived biomarkers HETE-CysPro and HETE-CysProPhe (15.6 nM SM in serum).Stability of the alkylated Met 329 in vitro and in vivo was limited to 5 days making this modification a beneficial short-time biomarker. Furthermore, it was found that the HETE-methionyl sulfonium ion can transfer its HETE moiety to the side chain of cysteine and glutamic acid as well as to the N-terminus of peptides and proteins in vitro thus revealing novel insights into the molecular toxicity of SM.

show abstract

Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

See 2 more Smart Citations

Methionine³²⁹ in human serum albumin: A novel target for alkylation by sulfur mustard

Siegert

Gandor

Kranawetvogl

et al. 2019

Drug Testing and Analysis

View full text Add to dashboard Cite

show abstract

“…Finally, it may be expected that the sample preparation will be more or less automated. Recently, a promising result was published by Carol-Visser et al (2008), who described the digestion and analysis of sulfur mustard adducts in albumin and the sarin adduct to BuChE.…”

Section: Concluding Remarks and Future Directionsmentioning

confidence: 99%

Laboratory Analysis of Chemical Warfare Agents, Adducts, and Metabolites in Biomedical Samples

Schans¹

2015

Handbook of Toxicology of Chemical Warfare Agents

View full text Add to dashboard Cite

Methods for Retrospective Detection of Exposure to Toxic Scheduled Chemicals. Part B: Mass Spectrometric and Immunochemical Analysis of Covalent Adducts to Proteins and DNA

Noort¹,

Schans²,

Black³

2020

Encyclopedia of Analytical Chemistry

View full text Add to dashboard Cite

In this article, an overview is presented of the methods that are currently available for retrospective detection of exposure to a number of chemical warfare agents (CWAs), based on adducts formed with macromolecules such as proteins. These methods can be applied for various purposes, e.g. diagnosis and dosimetry of exposure of casualties, confirmation of nonexposure, and verification of nonadherence to the Chemical Weapons Convention, health surveillance, and forensic purposes. The advantage of using protein adducts as biomarkers in comparison with free metabolites is that they are potentially much more long‐lived. The methods are predominantly based on liquid chromatography–mass spectrometry (LC–MS) analysis of enzymatic digests of the (modified) proteins or on selective removal of the specific adduct moiety from the protein, followed by gas chromatography–mass spectrometry (GC–MS) or LC–MS. Several of the methods have been successfully applied to actual cases and were shown to be highly retrospective.

show abstract

Development of an automated on-line pepsin digestion–liquid chromatography–tandem mass spectrometry configuration for the rapid analysis of protein adducts of chemical warfare agents

Cited by 45 publications

References 23 publications

Methionine³²⁹ in human serum albumin: A novel target for alkylation by sulfur mustard

Methionine³²⁹ in human serum albumin: A novel target for alkylation by sulfur mustard

Laboratory Analysis of Chemical Warfare Agents, Adducts, and Metabolites in Biomedical Samples

Methods for Retrospective Detection of Exposure to Toxic Scheduled Chemicals. Part B: Mass Spectrometric and Immunochemical Analysis of Covalent Adducts to Proteins and DNA

Contact Info

Product

Resources

About

Development of an automated on-line pepsin digestion–liquid chromatography–tandem mass spectrometry configuration for the rapid analysis of protein adducts of chemical warfare agents

Cited by 45 publications

References 23 publications

Methionine329 in human serum albumin: A novel target for alkylation by sulfur mustard

Methionine329 in human serum albumin: A novel target for alkylation by sulfur mustard

Laboratory Analysis of Chemical Warfare Agents, Adducts, and Metabolites in Biomedical Samples

Methods for Retrospective Detection of Exposure to Toxic Scheduled Chemicals. Part B: Mass Spectrometric and Immunochemical Analysis of Covalent Adducts to Proteins and DNA

Contact Info

Product

Resources

About

Methionine³²⁹ in human serum albumin: A novel target for alkylation by sulfur mustard

Methionine³²⁹ in human serum albumin: A novel target for alkylation by sulfur mustard