基于萘酰亚胺的生物硫醇探针的合成及对含硫醇氨基酸的检测

引用本文: 郭靖, 刘庆文, 杜建时, 孔祥怡, 宋岩, 杨清彪, 赵晴, 李耀先. 基于萘酰亚胺的生物硫醇探针的合成及对含硫醇氨基酸的检测[J]. 分析化学, 2017, 45(9): 1330-1338. doi: 10.11895/j.issn.0253-3820.170319 shu

Citation: GUO Jing, LIU Qing-Wen, DU Jian-Shi, KONG Xiang-Yi, SONG Yan, YANG Qing-Biao, ZHAO Qing, LI Yao-Xian. Synthesis of Naphthalimide-based Biothiols Probe and Detection of Amino Acids Containing Sulfhydryl Groups[J]. Chinese Journal of Analytical Chemistry, 2017, 45(9): 1330-1338. doi: 10.11895/j.issn.0253-3820.170319 shu

基于萘酰亚胺的生物硫醇探针的合成及对含硫醇氨基酸的检测

郭靖 ^1,2, ,
刘庆文 ^2, ,
杜建时 ^1, ,
孔祥怡 ^1, ,
宋岩 ^2, ,
杨清彪 ^2, ,
赵晴 ^1, ,
李耀先 ^2,

1.
吉林大学中日联谊医院, 长春 130031;2 吉林大学化学学院, 长春 130021
基金项目:
本文系吉林省自然科学基金（No.20160101311JC）和吉林省科技厅重点项目（Nos.20170204039GX，201307270299YY，20150204057SF）资助

摘要: 合成了以4-羟基萘酰亚胺为荧光团，2，4-二硝基苯磺酰氧基为特异性识别基团的生物硫醇探针4-（2，4-二硝基苯磺酰氧基）-正丁基-1，8-萘酰亚胺（DNSBN）。吸收光谱和荧光光谱结果表明，DNSBN对半胱氨酸（Cys）、同型半胱氨酸（Hcy）和谷胱甘肽（GSH）3种生物硫醇分子具有高效的检测识别能力，不受其它17种天然氨基酸的干扰。同时，通过荧光滴定实验证实了此探针是一种比率型探针，555 nm处的荧光强度与溶液中的生物硫醇分子浓度在0~20 μmol/L范围内呈良好的线性关系，对Cys、Hcy和GSH的检出限（3σ）分别为25.9、92.0和77.9 nmol/L。而吸收光谱、荧光光谱和质谱表征数据显示，生物硫醇与2，4-二硝基苯磺酸酯发生亲核取代反应并导致磺酸酯的分解。随着识别基团的解离，探针分子的d-PeT（donor-excited photoinduced electron transfer）效应被解除，并出现非常明显的比色与荧光变化。HeLa细胞成像实验表明，探针DNSBN具有良好的生物相容性，能够对细胞外源性生物硫醇分子进行检测。

关键词:

English

Synthesis of Naphthalimide-based Biothiols Probe and Detection of Amino Acids Containing Sulfhydryl Groups

1.
Department of Neurology, China-Japan Union Hospital of Jilin University, Changchun 130033, China;2 College of Chemistry, Jilin University, Changchun 130021, China
Received Date: 19 May 2017
Revised Date: 20 July 2017

Abstract: A novel probe (DNSBN) towards biothiols on the basis of 4-hydroxynaphthalimide as fluorophores and 2, 4-dinitrobenzenesulfonyloxy group as specific recognition site was designed and synthesized. The result of absorption and fluorescence spectral analyses indicated that the probe had high sensitivity and selectivity towards cysteine (Cys), homocysteine (Hcy) and glutathione (GSH), and the detection was not affected by other 17 kinds of natural amino acids. Meanwhile, it was confirmed that DNSBN was a ratiometric probe through the fluorescence titration experiment, and the fluorescent intensity at 555 nm had a high linear relationship with biothiols concentration in the range of 0-20 μmol/L. The detection limits (3σ) of Cys, Hcy and GSH were 25.9, 92.0 and 77.9 nmol/L, respectively. The absorption, emission and mass spectra indicated that biothiols could be engaged in nucleophilic substitution reaction with 2,4-dinitrobenzenesulfonate, which induced the sulfonic esters decomposed. With the departure of receptor unit, the d-PeT progress (donor-excited photoinduced electron transfer) was blocked with an obvious colorimetric and fluorescence change. Finally, HeLa cell imaging experiments verified that DNSBN had good biocompatibility and could be used to detect exogenous biothiols.

Key words:

1. [1]
  Wang Y, Zheng J, Zhang Z, Yuan C, Fu D. Colloids Surf. A,2009,342(1-3):102-106
2. [2]
  Zhang N, Qu F, Luo H Q, Li N B. Biosens. Bioelectron.,2013,42:214-218
3. [3]
  Shahrokhian S. Anal. Chem.,2001,73(24):5972-5978
4. [4]
  Cattaneo M. J. Thromb. Haemost.,1999,81(2):165-176
5. [5]
  Jager A, Kostense P J, Nijpels G, Dekker M J, Heine J R, Bouter M L, Donker M J, Stehouwer A D. Arterioscler. Thromb. Vasc. Biol.,2001,21(1):74
6. [6]
  McLean R R, Jacques P F, Selhub J, Tucker L K, Samelson J E, Broe E K, Hannan T M, Cupples A L, Kiel P D. N. Engl. J. Med.,2004,350(20):2042-2049
7. [7]
  van Meurs J B J, Dhonukshe-Rutten R A M, Pluijm S M F, van der Klift M, Jonge R, Lindemans J, Groot C P G M L, Hofman A, Witteman C M J, van Leeuwen P T M J, Breteler M B M, Lips P, Pols A P H, Uitterlinden G A. N. Engl. J. Med.,2004,350(20):2033-2041
8. [8]
  Morris M S. Lancet,2003,2(7):425-428
9. [9]
  Smach M A, Jacob N, Golmard J L, Charfeddine B, Lammouchi T, Ben Othman L, Dridi H, Bennamou S, Limem K. Eur. Neurol.,2011,65(5):270-278
10. [10]
  Townsend D M, Tew K D, Tapiero H, Biomed. Pharmacother.,2003,57(3-4):145-155
11. [11]
  Ivanov A R, Nazimov I V, Baratova L A. J. Chromatogr. A,2000,870(1-2):433-442
12. [12]
  Ivanov A R, Nazimov I V, Baratova L A. J. Chromatogr. A,2000,895(1-2):157-166
13. [13]
  Nekrassova O, Lawrence N S, Compton R G. Talanta,2003,60(6):1085-1095
14. [14]
  Xu K, Zhang Y, Tang B, Laskin J, Roach J.P, Chen H. Anal. Chem.,2010,82(16):6926-6932
15. [15]
  Aléssio A C M, Annichino-Bizzacchi J M, Bydlowski S P, Eberlin N M, Vellasco P A, H ehr F N. Am. J. Med. Genet. A,2004,128A(3):256-260
16. [16]
  Guo Y, Shao S, Xu J, Shi Y, Jiang S. Tetrahedron Lett.,2004,45(34):6477-6480
17. [17]
  Ros-Lis J V, García B, Jiménez D, Martínez-Máñez R, Sancenón F, Soto J, Gonzalvo F, Valldecabres C M. J. Am. Chem. Soc.,2004,126(13):4064-4065
18. [18]
  Zhu J, Dhimitruka I, Pei D. Org. Lett.,2004, 6(21):3809-3812
19. [19]
  Lee T P, Thomson E J, Karina A, Salter C, Johnston C, Davies G S, Compton G R. Analyst,2015,140(1):236-242
20. [20]
  Kim G J, Lee K, Kwon H, Kim H. Org. Lett.,2011,13(11):2799-2801
21. [21]
  Rusin O, Luce N N, Agbaria A R, Escobedo O J, Jiang S, Warner M I, Dawan B F, Lian K, Strongin M R. J. Am. Chem. Soc.,2004,126(2):438-439
22. [22]
  Lee J H, Lim C S, Tian Y S, Han J H, Cho B R. J. Am. Chem. Soc.,2010,132(4):1216-1217
23. [23]
  Shi Y, Yao J, Duan Y, Mi Q, Chen J, Xu Q, Gou G, Zhou Y, Zhang J. Bioorg. Med. Chem. Lett.,2013,23(9):2538-2542
24. [24]
  FU Xin, GU Dan-Yu, ZHAO Sheng-Dong, WEN Shi-Tong, ZHANG He. Chinese J. Anal. Chem.,2016,44(10):1487-1494傅昕, 顾丹玉, 赵圣东, 温世彤, 张何.分析化学,2016,44(10):1487-1494
25. [25]
  Wei M, Yin P, Shen Y, Zhang L, Deng J, Xue S, Li H, Guo B, Zhang Y, Yao S. Chem. Comm.,2013,49(41):4640-4642
26. [26]
  Zhang W, Huo F, Liu T, Yin C. Dyes Pigments,2016,133:248-254
27. [27]
  Liu T, Zhang X, Qiao Q, Zou C, Feng L, Cui J, Xu Z. Dyes Pigments,2013,99 (3):537-542
28. [28]
  Wei X, Yang X, Feng Y, Ning P, Yu H, Zhu M, Xi X, Guo Q, Meng X. Sens. Actuators B,2016,231:285-292
29. [29]
  Liao Y,Venkatesan P, Wei L, Wu S. Sens. Actuators B,2016,232:732-737
30. [30]
  Dai X, Wu Q, Wang P, Tian J, Xu Y, Wang S. Biosens. Bioelectron.,2014,59:35-39
31. [31]
  Sun J, Yang F, Zhao D, Chen C, Yang X. ACS Appl. Mater. Interfaces, 2015,7 (12):6860-6866
32. [32]
  WANG Xue, YANG Kun-Cheng, MAO Zhi-Yuan, HUANG Cheng-Zhi, WANG Jian. Chinese J. Anal. Chem.,2016,44(10):1482-1486王雪, 杨焜诚, 毛智远, 黄承志, 王健.分析化学,2016,44(10):1482-1486
33. [33]
  Yuan L, Lin W, Zhao S, Gao W, Chen B, He L, Zhu S. J. Am. Chem. Soc.,2012,134(32):13510-13523
34. [34]
  Schafer F Q, Buettner G R. J. Free Radicals Biol. Med.,2001,30(11):1191-1212
35. [35]
  Huo F, Kang J, Yin C, Zhang Y, Chao J. Sens. Actuators B,2015,207:139-143
36. [36]
  Yin J, Kwon Y, Kim D, Lee D, Kim G, Hu Y, Ryu J, Yoon J. J. Am. Chem. Soc.,2014,136:5351-5358

扫一扫看文章

计量

PDF下载量: 4
文章访问数: 605
HTML全文浏览量: 53

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

基于萘酰亚胺的生物硫醇探针的合成及对含硫醇氨基酸的检测

English

Synthesis of Naphthalimide-based Biothiols Probe and Detection of Amino Acids Containing Sulfhydryl Groups

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？

计量

通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

基于萘酰亚胺的生物硫醇探针的合成及对含硫醇氨基酸的检测

English

Synthesis of Naphthalimide-based Biothiols Probe and Detection of Amino Acids Containing Sulfhydryl Groups

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？

计量

文章相关

通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

CCS Chemistry：颜徐州、鲍哲南：你的皮肤可能是一片SPMs