一类新芥子气损伤生物标志物的体外高分辨质谱鉴定研究

引用本文: 杨展, 徐斌, 陈佳, 吴剑峰, 何跃忠, 谢剑炜. 一类新芥子气损伤生物标志物的体外高分辨质谱鉴定研究[J]. 分析化学, 2021, 49(10): 1733-1742. doi: 10.19756/j.issn.0253-3820.211256 shu

Citation: YANG Zhan, XU Bin, CHEN Jia, WU Jian-Feng, HE Yue-Zhong, XIE Jian-Wei. Identification of Some Novel Damage Biomarkers of Sulfur Mustard by High Resolution Mass Spectrometry in Vitro[J]. Chinese Journal of Analytical Chemistry, 2021, 49(10): 1733-1742. doi: 10.19756/j.issn.0253-3820.211256 shu

一类新芥子气损伤生物标志物的体外高分辨质谱鉴定研究

杨展 ^1,2, ,
徐斌 ^2, ,
陈佳 ^2, ,
吴剑峰 ^{2,
,} ,
何跃忠 ^1, ,
谢剑炜 ^2,

1.
军事科学院军事医学研究院, 北京 100850;
2.
军事科学院军事医学研究院毒物药物研究所, 抗毒药物与毒理学国家重点实验室, 北京 100850

通讯作者: 吴剑峰,E-mail:ammswjf@163.com

基金项目:
国家重点研发计划项目（Nos.2018YFC1602600，2020YFF0305000）资助。

摘要: 芥子气（SM）具有亲电性，是典型的高反应活性烷基化毒剂，人体经其暴露后，体内会生成多种水解氧化代谢产物和蛋白质、核酸加合物等，其中，二乙烯砜（Divinyl Sulfone，DVS）是近期发现的一种重要氧化代谢产物，具有类芥子气的反应活性和更高的毒性。以DVS及其谷胱甘肽（Glutathione，GSH）的二相代谢产物DVS-GSH分别染毒人血清白蛋白（HSA）与人源血浆，经蛋白酶K酶解和HLB小柱固相萃取净化后，采用UPLC-Q-Exactive Orbitrap高分辨质谱分析检测，鉴定出3种全新结构的白蛋白加合物：DVS-Cys-Pro-Phe （DVS-CPF）、Phe-Pro-Cys-DVS-Cys-Pro-Phe （FPC-DVS-CPF）和GSH-DVS-Cys-Pro-Phe （GSH-DVS-CPF）。研究结果表明，含有高反应活性烯键的DVS分子可与白蛋白的Cys-34残基发生特异性加合反应，并且其二相代谢产物DVS-GSH仍可继续发生加合反应，形成一类新的白蛋白加合物。鉴定的新结构加合物不仅为确证SM暴露和中毒诊断提供了新的损伤生物标志物，也为从新的角度阐述SM的损伤机制提供了实验证据。

关键词:

English

Identification of Some Novel Damage Biomarkers of Sulfur Mustard by High Resolution Mass Spectrometry in Vitro

1.
Academy of Military Medical Sciences, Academy of Military Sciences, Beijing 100850, China;
2.
State Key Laboratory of Toxicology and Medical Countermeasures and Laboratory of Toxicant Analysis, Institute of Pharmacology and Toxicology, Academy of Military Medical Sciences, Beijing 100850, China

Corresponding author: WU Jian-Feng, ammswjf@163.com

Received Date: 28 March 2021
Revised Date: 02 June 2021
Fund Project:
Supported by the National Key Research and Development Program of China(Nos.2018YFC1602600, 2020YFF0305000).

Abstract: Sulfur mustard (SM) is a typically representative alkylating agent with high reactivity. SM can produce a variety of hydrolytic and oxidative metabolites including various proteins and nucleic acid adducts. Among them, divinyl sulfone (DVS) is an important oxidative metabolite that should be paid more attention, which has high reactivity and toxicity close to SM. In this study, the human serum albumin (HSA) and human plasma were exposed to DVS and its two-phase metabolite DVS-GSH, respectively. After digestion with proteinase K and purification by solid phase extraction (SPE), UPLC-Q-extractive orbitrap high rosolution mass spectrometry (HRMS) was used to identify some albumin adducts. According to the analysis results, three novel biomarkers, DVS-Cys-Pro-Phe, Phe-Pro-Cys-DVS-Cys-Pro-Phe and GSH-DVS-Cys-Pro-Phe, were successfully identified by UPLC-Q-exactive orbitrap/HRMS. The results showed that both DVS and DVS-GSH could react with Cys-34 of albumin due to their highly reactive alkene bonds. These novel damage biomarkers not only provided new clues for SM exposure and diagnosis, but also provided evidence for elucidating the damage mechanism of SM from a new perspective.

Key words:

Sulfur mustard
/ Divinyl sulfone
/ Albumin adducts
/ Ultra performance liquid chromatography-tandem mass spectrometry

1. [1]
  PANAHI Y, ABDOLGHAFFARI A H, SAHEBKAR A. J. Cell. Biochem., 2018, 119(1):197-206.PANAHI Y, ABDOLGHAFFARI A H, SAHEBKAR A. J. Cell. Biochem., 2018, 119(1):197-206.
2. [2]
  TAEBI G, SOROUSH M, MODIRIAN E, KHATERI S, MOUSAVI B Z. Ganjparvar. Iran. J. War Public Health, 2015, 7(2):115-121.TAEBI G, SOROUSH M, MODIRIAN E, KHATERI S, MOUSAVI B Z. Ganjparvar. Iran. J. War Public Health, 2015, 7(2):115-121.
3. [3]
  JIANG A, MAIBACH H. J. Appl. Toxicol., 2018, 38(1):108-112.JIANG A, MAIBACH H. J. Appl. Toxicol., 2018, 38(1):108-112.
4. [4]
  CZUB M, NAWALA J, POPIEL S, DZIEDZIC D, BRZEZINSKI T, MASZCZYK P, SANDERSON H, FABISIAK J, BELDOWSKI J, KOTWICKI L. Mar. Environ. Res., 2020, 161:105077.CZUB M, NAWALA J, POPIEL S, DZIEDZIC D, BRZEZINSKI T, MASZCZYK P, SANDERSON H, FABISIAK J, BELDOWSKI J, KOTWICKI L. Mar. Environ. Res., 2020, 161:105077.
5. [5]
  QI M, XU B, WU J, ZHANG Y, ZONG C, CHEN J, GUO L, XIE J. J. Chromatogr. B:Anal. Technol. Biomed. Life Sci., 2016, 1028:42-50.QI M, XU B, WU J, ZHANG Y, ZONG C, CHEN J, GUO L, XIE J. J. Chromatogr. B:Anal. Technol. Biomed. Life Sci., 2016, 1028:42-50.
6. [6]
  OHEIX E, GRAVEL E, DORIS E. Chemistry, 2021, 27(1):54-68.OHEIX E, GRAVEL E, DORIS E. Chemistry, 2021, 27(1):54-68.
7. [7]
  LV S, ZHANG Y, XU B, XU H, ZHAO Y, CHEN J, GAO Z, WU J, XIE J. Chem. Res. Toxicol., 2017, 30(10):1874-1882.LV S, ZHANG Y, XU B, XU H, ZHAO Y, CHEN J, GAO Z, WU J, XIE J. Chem. Res. Toxicol., 2017, 30(10):1874-1882.
8. [8]
  POPIEL S, NAWAŁA J, DZIEDZIC D, GORDON D, DAWIDZIUK B. Int. J. Chem. Kinet., 2018, 50(2):75-89.POPIEL S, NAWAŁA J, DZIEDZIC D, GORDON D, DAWIDZIUK B. Int. J. Chem. Kinet., 2018, 50(2):75-89.
9. [9]
  FORD-MOORE A H. J. Chem. Soc., 1949, 512:2433-2440.FORD-MOORE A H. J. Chem. Soc., 1949, 512:2433-2440.
10. [10]
  WEST J, STAMM C E, BROWN H A, JUSTICE S L, MORANO K A. Chem. Res. Toxicol., 2011, 24(9):1457-1459.WEST J, STAMM C E, BROWN H A, JUSTICE S L, MORANO K A. Chem. Res. Toxicol., 2011, 24(9):1457-1459.
11. [11]
  WANG P, ZHANG Y, CHEN J, GUO L, XU B, WANG L, XU H, XIE J. Chem. Res. Toxicol., 2015, 28(6):1224-1233.WANG P, ZHANG Y, CHEN J, GUO L, XU B, WANG L, XU H, XIE J. Chem. Res. Toxicol., 2015, 28(6):1224-1233.
12. [12]
  ANSLOW W P, KARNOFSKY D A, VAL JAGER B, SMITH H W. J. Pharmacol. Exp. Ther., 1948, 93(1):1-9.ANSLOW W P, KARNOFSKY D A, VAL JAGER B, SMITH H W. J. Pharmacol. Exp. Ther., 1948, 93(1):1-9.
13. [13]
  SANTA CRUZ BIOTECHNOLOGY Inc. Divinyl Sulfone (MSDS no. sc-255120). In:Biotechnology S C ed. Safty Data Sheeted. Heidelberg:Germany, 2017:7.SANTA CRUZ BIOTECHNOLOGY Inc. Divinyl Sulfone (MSDS no. sc-255120). In:Biotechnology S C ed. Safty Data Sheeted. Heidelberg:Germany, 2017:7.
14. [14]
  DOS SANTOS J C S, RUEDA N, BARBOSA O, MILLÁN-LINARES M D C, PEDROCHE J, YUSTE M D, GONÇALVES L R B, FERNANDEZ-LAFUENTE R. J. Mol. Catal. B:Enzym., 2015, 117:38-44.DOS SANTOS J C S, RUEDA N, BARBOSA O, MILLÁN-LINARES M D C, PEDROCHE J, YUSTE M D, GONÇALVES L R B, FERNANDEZ-LAFUENTE R. J. Mol. Catal. B:Enzym., 2015, 117:38-44.
15. [15]
  DOS SANTOS J C S, RUEDA N, TORRES R, BARBOSA O, GONÇALVES L R B, FERNANDEZ-LAFUENTE R. Process Biochem., 2015, 50(6):918-927.DOS SANTOS J C S, RUEDA N, TORRES R, BARBOSA O, GONÇALVES L R B, FERNANDEZ-LAFUENTE R. Process Biochem., 2015, 50(6):918-927.
16. [16]
  MORALES-SANFRUTOS J, LOPEZ-JARAMILLO J, ORTEGA-MUÑOZ M, MEGIA-FERNANDEZ A, PEREZ-BALDERAS F, HERNANDEZ-MATEO F, SANTOYO-GONZALEZ F. Org. Biomol. Chem., 2010, 8(3):667-675.MORALES-SANFRUTOS J, LOPEZ-JARAMILLO J, ORTEGA-MUÑOZ M, MEGIA-FERNANDEZ A, PEREZ-BALDERAS F, HERNANDEZ-MATEO F, SANTOYO-GONZALEZ F. Org. Biomol. Chem., 2010, 8(3):667-675.
17. [17]
  BELLAART A C. Phosphorus Sulfur Relat. Elem., 2007, 6(1-2):33-34.BELLAART A C. Phosphorus Sulfur Relat. Elem., 2007, 6(1-2):33-34.
18. [18]
  CHENG X, LIU C, YANG Y, LIANG L, CHEN B, YU H, XIA J, LIU S, LI Y. Toxicol. Lett., 2021, 344:46-57.CHENG X, LIU C, YANG Y, LIANG L, CHEN B, YU H, XIA J, LIU S, LI Y. Toxicol. Lett., 2021, 344:46-57.
19. [19]
  JOHN H, WILLOH S, HORMANN P, SIEGERT M, VONDRAN A, THIERMANN H. Anal. Chem., 2016, 88(17):8787-8794.JOHN H, WILLOH S, HORMANN P, SIEGERT M, VONDRAN A, THIERMANN H. Anal. Chem., 2016, 88(17):8787-8794.
20. [20]
  JOHN H, SIEGERT M, GANDOR F, GAWLIK M, KRANAWETVOGL A, KARAGHIOSOFF K, THIERMANN H. Toxicol. Lett., 2016, 244:103-111.JOHN H, SIEGERT M, GANDOR F, GAWLIK M, KRANAWETVOGL A, KARAGHIOSOFF K, THIERMANN H. Toxicol. Lett., 2016, 244:103-111.
21. [21]
  PANTAZIDES B G. CROW B S, GARTON J W, QUINONES-GONZALEZ J A, BLAKE T A, THOMAS J D, JOHNSON R C. Chem. Res. Toxicol., 2015, 28(2):256-261.PANTAZIDES B G. CROW B S, GARTON J W, QUINONES-GONZALEZ J A, BLAKE T A, THOMAS J D, JOHNSON R C. Chem. Res. Toxicol., 2015, 28(2):256-261.
22. [22]
  LIU C, LIANG L, XIANG Y, YU H, ZHOU S, XI H, LIU S, LIU J. Anal. Bioanal. Chem., 2015, 407(23):7037-7046.LIU C, LIANG L, XIANG Y, YU H, ZHOU S, XI H, LIU S, LIU J. Anal. Bioanal. Chem., 2015, 407(23):7037-7046.
23. [23]
  GANDOR F, GAWLIK M, THIERMANN H, JOHN H. J. Anal. Toxicol., 2015, 39(4):270-279.GANDOR F, GAWLIK M, THIERMANN H, JOHN H. J. Anal. Toxicol., 2015, 39(4):270-279.
24. [24]
  LIANG Long-Hui, LIU Shi-Lei, LIU Chang-Cai, HUANG Gui-Lan, XIANG Yu, LI Xin-Hai, YU Hui-Lan. Chin. J. Anal. Chem., 2016, 44(11):1763-1770. 梁龙辉, 刘石磊, 刘昌财, 黄桂兰, 向宇, 李欣海, 于惠兰. 分析化学, 2016, 44(11):1763-1770.
25. [25]
  NOORT D, HULST A G, JANSEN R. Arch. Toxicol., 2002, 76(2):83-88.NOORT D, HULST A G, JANSEN R. Arch. Toxicol., 2002, 76(2):83-88.
26. [26]
  OWEN J B, BUTTERFIELD D A. Methods Mol. Biol., 2010, 648:269-277.OWEN J B, BUTTERFIELD D A. Methods Mol. Biol., 2010, 648:269-277.
27. [27]
  YU Y, CHAU Y. Biomacromolecules, 2012, 13(3):937-942.YU Y, CHAU Y. Biomacromolecules, 2012, 13(3):937-942.

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

一类新芥子气损伤生物标志物的体外高分辨质谱鉴定研究

通讯作者: 吴剑峰,E-mail:ammswjf@163.com

English

Identification of Some Novel Damage Biomarkers of Sulfur Mustard by High Resolution Mass Spectrometry in Vitro

Corresponding author: WU Jian-Feng, ammswjf@163.com

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通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

一类新芥子气损伤生物标志物的体外高分辨质谱鉴定研究

通讯作者: 吴剑峰,E-mail:ammswjf@163.com

English

Identification of Some Novel Damage Biomarkers of Sulfur Mustard by High Resolution Mass Spectrometry in Vitro

Corresponding author: WU Jian-Feng, ammswjf@163.com

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南： 你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

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通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

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