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Title: Study of Highly Selective and Efficient Thiol Derivatization using Selenium Reagents by Mass Spectrometry

Abstract

Biological thiols are critical physiological components and their detection often involves derivatization. This paper reports a systemic mass spectrometry (MS) investigation of the cleavage of Se-N bond by thiol to form a new Se-S bond, the new selenium chemistry for thiol labeling. Our data shows that the reaction is highly selective, rapid, reversible and efficient. For instance, among twenty amino acids, only cysteine was found to be reactive with Se-N containing reagents and the reaction takes place in seconds. By adding dithiothreitol (DTT), the newly formed Se-S bond of peptides/proteins can be reduced back to free thiol. The high selectivity and excellent reversibility of the reaction provide potential of using this chemistry for selective identification of thiol compounds or enriching and purifying thiol peptides/proteins. In addition, the derivatized thiol peptides have interesting dissociation behavior, which is tunable using different selenium reagents. For example, by introducing an adjacent nucleophilic group into the selenium reagent in the case of using ebselen, the reaction product of ebselen with glutathione (GSH) is easy to lose the selenium tag upon collision-induced dissociation (CID), which is useful to "fish out" those peptides containing free cysteine residues by precursor ion scan. By contrast, the selenium tag ofmore » N-(phenylseleno) phthalimide reagent can be stable and survive in CID process, which would be of value in pinpointing thiol location using a top-down proteomic approach. Also, the high conversion yield of the reaction allows the counting of total number of thiol in proteins. We believe that ebselen or N-(phenylseleno) phthalimide as tagging thiol-protein reagents will have important applications in both qualitative and quantitative analysis of different thiol-proteins derived from living cells by MS method.« less

Authors:
; ; ; ; ;
Publication Date:
Research Org.:
Pacific Northwest National Laboratory (PNNL), Richland, WA (US), Environmental Molecular Sciences Laboratory (EMSL)
Sponsoring Org.:
USDOE
OSTI Identifier:
988657
Report Number(s):
PNNL-SA-72020
34920; KC0302020; TRN: US201018%%610
DOE Contract Number:  
AC05-76RL01830
Resource Type:
Journal Article
Journal Name:
Analytical Chemistry, 82(16):6926-6932
Additional Journal Information:
Journal Volume: 82; Journal Issue: 16
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 99 GENERAL AND MISCELLANEOUS//MATHEMATICS, COMPUTING, AND INFORMATION SCIENCE; AMINO ACIDS; CHEMISTRY; CLEAVAGE; CYSTEINE; DERIVATIZATION; DETECTION; DISSOCIATION; GLUTATHIONE; MASS SPECTROSCOPY; PEPTIDES; PRECURSOR; PROTEINS; RESIDUES; SELENIUM; THIOLS; mass spectrometry; thiol derivatization; selenium reagent; peptide/protein; reaction mechanism; Environmental Molecular Sciences Laboratory

Citation Formats

Xu, Kehua, Zhang, Yun W, Tang, Bo, Laskin, Julia, Roach, Patrick J, and Chen, Hao. Study of Highly Selective and Efficient Thiol Derivatization using Selenium Reagents by Mass Spectrometry. United States: N. p., 2010. Web. doi:10.1021/ac1011602.
Xu, Kehua, Zhang, Yun W, Tang, Bo, Laskin, Julia, Roach, Patrick J, & Chen, Hao. Study of Highly Selective and Efficient Thiol Derivatization using Selenium Reagents by Mass Spectrometry. United States. doi:10.1021/ac1011602.
Xu, Kehua, Zhang, Yun W, Tang, Bo, Laskin, Julia, Roach, Patrick J, and Chen, Hao. Sun . "Study of Highly Selective and Efficient Thiol Derivatization using Selenium Reagents by Mass Spectrometry". United States. doi:10.1021/ac1011602.
@article{osti_988657,
title = {Study of Highly Selective and Efficient Thiol Derivatization using Selenium Reagents by Mass Spectrometry},
author = {Xu, Kehua and Zhang, Yun W and Tang, Bo and Laskin, Julia and Roach, Patrick J and Chen, Hao},
abstractNote = {Biological thiols are critical physiological components and their detection often involves derivatization. This paper reports a systemic mass spectrometry (MS) investigation of the cleavage of Se-N bond by thiol to form a new Se-S bond, the new selenium chemistry for thiol labeling. Our data shows that the reaction is highly selective, rapid, reversible and efficient. For instance, among twenty amino acids, only cysteine was found to be reactive with Se-N containing reagents and the reaction takes place in seconds. By adding dithiothreitol (DTT), the newly formed Se-S bond of peptides/proteins can be reduced back to free thiol. The high selectivity and excellent reversibility of the reaction provide potential of using this chemistry for selective identification of thiol compounds or enriching and purifying thiol peptides/proteins. In addition, the derivatized thiol peptides have interesting dissociation behavior, which is tunable using different selenium reagents. For example, by introducing an adjacent nucleophilic group into the selenium reagent in the case of using ebselen, the reaction product of ebselen with glutathione (GSH) is easy to lose the selenium tag upon collision-induced dissociation (CID), which is useful to "fish out" those peptides containing free cysteine residues by precursor ion scan. By contrast, the selenium tag of N-(phenylseleno) phthalimide reagent can be stable and survive in CID process, which would be of value in pinpointing thiol location using a top-down proteomic approach. Also, the high conversion yield of the reaction allows the counting of total number of thiol in proteins. We believe that ebselen or N-(phenylseleno) phthalimide as tagging thiol-protein reagents will have important applications in both qualitative and quantitative analysis of different thiol-proteins derived from living cells by MS method.},
doi = {10.1021/ac1011602},
journal = {Analytical Chemistry, 82(16):6926-6932},
number = 16,
volume = 82,
place = {United States},
year = {2010},
month = {8}
}