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Identification of creatine kinase and alpha‐1 antitrypsin as protein targets of alkylation by sulfur mustard
- Source :
- Drug Testing and Analysis. 13:268-282
- Publication Year :
- 2020
- Publisher :
- Wiley, 2020.
-
Abstract
- 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.
- Subjects :
- Models, Molecular
Alkylation
Pharmaceutical Science
Mass spectrometry
Proteomics
01 natural sciences
Analytical Chemistry
03 medical and health sciences
chemistry.chemical_compound
0302 clinical medicine
Mustard Gas
medicine
Humans
Environmental Chemistry
Chemical Warfare Agents
030216 legal & forensic medicine
Creatine Kinase
Spectroscopy
chemistry.chemical_classification
Gel electrophoresis
010401 analytical chemistry
Sulfur mustard
Human serum albumin
0104 chemical sciences
Amino acid
HEK293 Cells
chemistry
Biochemistry
alpha 1-Antitrypsin
medicine.drug
Cysteine
Subjects
Details
- ISSN :
- 19427611 and 19427603
- Volume :
- 13
- Database :
- OpenAIRE
- Journal :
- Drug Testing and Analysis
- Accession number :
- edsair.doi.dedup.....a90f190e5423e97b4bde0d2c938b981c