成分分析

基于罗丹明B为荧光基团的三肽荧光探针的设计及应用研究*

展开
  • 1.青海民族大学药学院,西宁 810007;
    2.青海省青藏高原植物化学重点实验室,西宁 810007;
    3.青海省植物药物分析重点实验室,西宁 810007
第一作者 Tel:18300666252;E-mail:guobingxve@163.com
**Tel:15293111864;E-mail:wujiang@lzu.edu.cn

修回日期: 2023-02-28

  网络出版日期: 2024-06-25

基金资助

*青海省创新平台建设项目(2022-ZJ-Y14);青海省药材资源保护与高值化利用创新团队(2021XJPI02)

Design and application of tripeptide fluorescent probe based on rhodamine B as fluorophore*

Expand
  • 1. College of Pharmacy,Qinghai Nationalities University, Xining 810007, China;
    2. Key Laboratory of Plant Resources of Qinghai-Tibet Plateau in Chemical Research, Xining 810007, China;
    3. Key Laboratory of Pharmaceutical Analysis in Qinghai Province, Xining 810007, China

Revised date: 2023-02-28

  Online published: 2024-06-25

摘要

目的: 基于铜离子和硫离子对罗丹明B标记的多肽荧光猝灭和荧光恢复作用,建立在活细胞和超纯水中连续检测Cu2+和S2-的方法。方法: 采用多肽固相合成技术制备了RhB-Gly-Gly-His,它与Cu2+等量结合形成配合物,该配合物可检测活细胞中的S2-、半胱氨酸(Cys)、谷光甘肽(GSH),同时通过Job曲线和计算机辅助模拟计算预测了该荧光传感器的结构和结合模式。结果: 该多肽荧光传感器能与Cu2+等量结合发生荧光淬灭,使用LOD=3σ/k方程计算的荧光滴定测量的检测限为143 nmol·L-1 (Cu2+)和408 nmol·L-1 (S2-)。结论: 成功地合成了RhB-Gly-Gly-His,该荧光传感器具有荧光发射强、水溶性好、生物相容性好、毒性低等特点。该荧光传感器具有较高的选择性和灵敏度,已成功地用于Cu2+和S2-的定性和定量检测,且拥有良好的选择性,在细胞中的体内成像突出了其在S2-,Cys,GSH相关疾病的病理诊断中具有广阔的前景。

本文引用格式

郭冰雪, 吴疆, 林鹏程, 王欢, 王敏, 穆文成, 任益萱, 周蒋, 苏敬蔚 . 基于罗丹明B为荧光基团的三肽荧光探针的设计及应用研究*[J]. 药物分析杂志, 2023 , 43(6) : 930 -938 . DOI: 10.16155/j.0254-1793.2023.06.03

Abstract

Objective: To establish a method for the continuous detection of Cu2+ and S2- in living cells and ultrapure water based on the fluorescence bursting and fluorescence recovery effects of Cu2+ and S2- on rhodamine B-labeled peptides. Methods: RhB-Gly-Gly-His was prepared by solid-phase peptide synthesis technique. RhB-Gly-Gly-His was equivalently bound with Cu2+ to form a complex which could detect S2-, Cys, and GSH in living cells. The structure and binding mode of this fluorescent sensor were predicted by Job curve and computer-aided simulation calculation. Results: The peptide fluorescent sensor binds with Cu2+ in equal amounts to undergo fluorescence quenching. The detection limits of fluorescence titration measurements were 143 nmol·L-1 (Cu2+) and 408 nmol·L-1 (S2-) with the LOD=3σ/k equation. Conclusion: RhB-Gly-Gly- His was successfully synthesized, and the fluorescent sensor has strong fluorescence emission, good water solubility, good biocompatibility and low toxicity. With high selectivity and sensitivity, this fluorescent sensor has been successfully used for the qualitative and quantitative detection of Cu2+ and S2-. This fluorescent sensor possesses good selectivity for in vivo imaging in cells and highlights a promising future in the pathological diagnosis of S2-, Cys, and GSH in related diseases.

参考文献

[1] AN Y, WANG P, YUE Z. A sequential and reversibility fluorescent pentapeptide probe for Cu(Ⅱ) ions and hydrogen sulfide detections and its application in two different living cells imaging[J].Spectrochim Acta A Mol Biomol Spectrosc, 2019, 216:319
[2] WANG P, WU J, SU P, et al. Fluorescence “on-off-on” peptide-based chemosensor for the selective detection of Cu2+ and S2- and its application in living cell bioimaging[J].Dalton Trans, 2016, 45(41):16246
[3] WANG P, WU J, ZHAO C. A water-soluble peptide fluorescent chemosensor for detection of cadmium(Ⅱ) and copper(Ⅱ) by two different response modes and its application in living LNcap cells[J].Spectrochim Acta A Mol Biomol Spectrosc, 2020, 226:117600
[4] SONG RQ, MA X, SONG GY. Tb(Ⅲ)-based coordination polymer for detection of Cu(Ⅱ) ion and treatment effect on Alzheimer’s disease by reducing inflammatory response and inflammatory cytokines release[J].J Chin Chem Soc, 2020, 67(8):1462
[5] WANG P, XUE S, YANG X. Highly selective and sensitive detection of hydrogen sulfide in aqueous medium and live cells using peptide-based bioprobe to mimic the binding sites of the ceruloplasmin for Cu(II) ions[J].Biosens Bioelectron, 2020, 163:112283
[6] SHAO H, YIN D, LI D, et al. Simultaneous visual detection and removal of Cu(2+) with electrospun self-supporting flexible amidated polyacrylonitrile/branched polyethyleneimine nanofiber membranes[J].ACS Appl Mater Interfaces, 2021, 13(41):49288
[7] GAETKE L M, CHOW C K. Copper toxicity, oxidative stress, and antioxidant nutrients[J].Toxicology, 2003, 189(1-2):147
[8] XU J, LIU N, HAO C, et al. A novel fluorescence “on-off-on” peptide-based chemosensor for simultaneous detection of Cu2+, Ag+ and S2-[J].Sens Actuators B: Chem, 2019, 280:129
[9] WANG P, WU X, WU J, et al. Highly selective and sensitive peptide-based fluorescent chemosensor for detection of Zinc(Ⅱ) ions in aqueous medium and living cells[J].J Photochem Photobiol A: Chem, 2019, 382:11929
[10] WANG Q, GUO Z, ZHOU D, et al. A novel fluorescent probe for highly selective and sensitive detection of sulfur ions in real samples and living cells based on the tripeptide-Cu2+ ensemble system[J].Microchem J, 2021, 169:106612
[11] WANG P, SUN L, WU J, et al. A dual-functional colorimetric and fluorescent peptide-based probe for sequential detection of Cu(2+) and S(2-) in 100% aqueous buffered solutions and living cells[J].J Hazard Mater, 2021, 407:124388
[12] WANG P, ZHOU D, XUE S, et al. Rational design of dual-functional peptide-based chemosensor for sequential detection of Ag+(AgNPs) and S2- ions by fluorescent and colorimetric changes and its application in live cells, real water samples and test strips[J].Microchem J, 2022, 177:107236
[13] WANG P, DUAN L, LIAO Y. A retrievable and highly selective peptide-based fluorescent probe for detection of Cd2+ and Cys in aqueous solutions and live cells[J].Microchemical Journal, 2019, 146:818
[14] WANG P, WU J, SU P, et al. A novel fluorescent chemosensor based on tetra-peptides for detecting zinc ions in aqueous solutions and live cells[J].J Mater Chem B, 2016, 4(26):4526
[15] WANG P, WU J, AN Y, et al. A novel fluorescent chemosensor for detection of Zn(Ⅱ) ions based on dansyl-appended dipeptide in two different living cells[J].Spectrochim Acta A Mol Biomol Spectrosc, 2019, 220:117140
[16] WANG P, WANG S, CHEN L, et al. A novel peptide-based fluorescent probe for sensitive detection of zinc(Ⅱ) and its applicability in live cell imaging[J].Spectrochim Acta A Mol Biomol Spectrosc, 2020, 240:118549
[17] WANG P, AN Y, WU J. Highly sensitive turn-on detection of mercury(Ⅱ) in aqueous solutions and live cells with a chemosensor based on tyrosine[J].Spectrochim Acta A Mol Biomol Spectrosc, 2020, 230:118004
[18] LOHANI CR, NEUPANE LN, KIM JM, et al. Selectively and sensitively monitoring Hg2+ in aqueous buffer solutions with fluorescent sensors based on unnatural amino acids[J].Sens Actuat B:Chem, 2012, 161(1):1088
[19] WANG P, WU J, SU P, et al. Fluorescence “on-off-on” peptide-based chemosensor for the selective detection of Cu(2+) and S(2-) and its application in living cell bioimaging[J].Dalton Trans, 2016, 45(41):16246
[20] WANG P, ZHOU D, CHEN B. High selective and sensitive detection of Zn(Ⅱ) using tetrapeptide-based dansyl fluorescent chemosensor and its application in cell imaging[J].Spectrochim Acta A Mol Biomol Spectrosc, 2018, 204:735
[21] XUE S, WANG P, CHEN K. A novel fluorescent chemosensor for detection of mercury(Ⅱ) ions based on dansyl-peptide and its application in real water samples and living LNcap cells[J].Spectrochim Acta A Mol Biomol Spectrosc, 2020, 226:117616
[22] WANG P, ZHOU D, CHEN B. A fluorescent dansyl-based peptide probe for highly selective and sensitive detect Cd(2+) ions and its application in living cell imaging[J].Spectrochim Acta A Mol Biomol Spectrosc, 2019, 207:276
文章导航

/