硫代黄素T与溶菌酶淀粉样纤维结合的荧光停流动力学

doi:10.3969/j.issn.1000-1565.2017.05.006

河北大学学报(自然科学版) ›› 2017, Vol. 37 ›› Issue (5): 476-482.DOI: 10.3969/j.issn.1000-1565.2017.05.006

硫代黄素T与溶菌酶淀粉样纤维结合的荧光停流动力学

马刚, 秦哲,刘思阳,商艳丽,贾宝环

收稿日期:2017-03-26 出版日期:2017-09-25 发布日期:2017-09-25
通讯作者: 马刚(1971—),男,北京人,河北大学教授,博士,主要从事与蛋白纤维化聚集相关的研究. E-mail:gangma@hbu.edu.cn;商艳丽(1979—),女,河北保定人,河北大学副教授,博士,主要从事材料化学方面的研究.E-mail:ylshang@hbu.edu.cn
作者简介:马刚(1971—),男,北京人,河北大学教授,博士,主要从事与蛋白纤维化聚集相关的研究. E-mail:gangma@hbu.edu.cn
基金资助:
国家自然科学基金资助项目(21075027);河北省自然科学基金资助项目(B2011201082;B2016201034)

Fluorescence-detected stopped-flow kinetic investigation of thioflavin T binding onto the amyloid fibril of lysozyme

MA Gang, QIN Zhe, LIU Siyang, SHANG Yanli, JIA Baohuan

Key Laboratory of Medicinal Chemistry and Molecular Diagnosis of Ministry of Education, College of Chemistry and Environmental Science, Hebei University, Baoding 071002, China

Received:2017-03-26 Online:2017-09-25 Published:2017-09-25

摘要/Abstract

摘要： 蛋白淀粉样纤维与多种疾病有关,如阿尔茨海默病、帕金森病、Ⅱ型糖尿病等.荧光染料硫代黄素T(ThT)广泛地应用于淀粉样纤维的定性和定量测定.但从目前的研究来看,ThT与淀粉样纤维结合的动力学机理研究尚非常少见.本研究应用荧光停流动力学技术对盐酸胍诱导的溶菌酶淀粉样纤维和ThT的结合动力学进行了研究,提出ThT与溶菌酶淀粉样纤维结合的动力学符合双位点平行反应机理模型.本研究对深入理解荧光染料分子与蛋白淀粉样纤维的作用有借鉴意义.

关键词: 硫代黄素T, 溶菌酶, 淀粉样纤维, 动力学

Abstract: Deposition of amyloid fibrils in tissues and organs is relevant to a wide range of devastating human diseases, including Alzheimer’s disease, Parkinson’s disease, and type Ⅱ diabetes. Thioflavin T(ThT)is widely used as a fluorescent probe for qualitative and quantitative detections of amyloid fibrils. Yet the exact kinetic mechanism of ThT binding onto amyloid fibril remains elusive. In this study, we used fluorescence-detected stopped-flow technique to study the binding mechanism of ThT onto the amyloid fibril of hen egg white lysozyme induced by guanidine hydrochloride. We proposed a kinetic binding mechanism to interpret our experimental observations. Our results showed that the binding of ThT onto lysozyme fibril follow a dual-site parallel binding mechanism. We believe our work is beneficial for better understanding of the interaction between fluorescent dye and amyloid fibril.

Key words: thioflavin T, lysozyme, amyloid fibril, kinetics

中图分类号:

O643

马刚, 秦哲,刘思阳,商艳丽,贾宝环. 硫代黄素T与溶菌酶淀粉样纤维结合的荧光停流动力学[J]. 河北大学学报(自然科学版), 2017, 37(5): 476-482.

MA Gang, QIN Zhe, LIU Siyang, SHANG Yanli, JIA Baohuan. Fluorescence-detected stopped-flow kinetic investigation of thioflavin T binding onto the amyloid fibril of lysozyme[J]. Journal of Hebei University (Natural Science Edition), 2017, 37(5): 476-482.

参考文献

[1] NELSON R, EISENBERG D. Structural models of amyloid‐like fibrils[J]. Advances in Protein Chemistry,2006, 73: 235-282. DOI:10.1016/S0065-3233(06)73008-X.
[2] SAWAYA M R, SAMBASHIVAN S, NELSON R, et al. Atomic structures of amyloid cross-beta spines reveal varied steric zippers[J]. Nature, 2007, 447: 453-457. DOI:10.1038/nature05695.
[3] CHITI F, DOBSON C M. Protein misfolding, functional amyloid, and human disease[J]. Annual Review of Biochemistry, 2006, 75: 333-366. DOI: 10.1146/annurev.biochem.75.101304.123901.
[4] LANSBURY P T, LASHUEL H A. A century-old debate on protein aggregation and neurodegeneration enters the clinic[J]. Nature, 2006, 443: 774-779. DOI:10.1038/nature05290.
[5] SACHSE C, FAENDRICH M, GRIGORIEFF N. Paired beta-sheet structure of an a beta(1-40)amyloid fibril revealed by electron microscopy[J]. Proceedings of the National Academy of Sciences of the United States of America, 2008, 105: 7462-7466. DOI: 10.1073/pnas.0712290105.
[6] TYCKO R, WICKNER R B. Molecular structures of amyloid and prion fibrils: Consensus versus controversy[J]. Accounts of Chemical Research, 2013, 46: 1487-1496. DOI: 10.1021/ar300282r.
[7] 张岚, 杨延莲, 王琛. 调节分子对茁淀粉样蛋白的聚集调控及细胞毒性抑制效应[J]. 生物物理学报, 2010, 26: 649-661. ZHANG L, YANG Y L, WANG C. Regulation of aggregation and cytotoxicity of β-Amyloid by molecular modulators[J]. ACTA Biophysica sinica, 2010,26:649-661.
[8] 刘鹏, 赵玉芬, 李艳梅. Tau蛋白介导阿尔兹海默病的机理及相关药物[J]. 中国科学:化学, 2010, 40: 906-913. LIU P, ZHAO Y F, LI Y M. Advances in Tau-mediated neurodegeneration and Tau-focused drug for Alzhehimer’s disease[J]. Scientia Sinica Chimica, 2010,40:906-913.
[9] 刘夫锋, 董晓燕, 孙彦. 海藻糖抑制淀粉质多肽42构象转变的分子动力学模拟[J]. 物理化学学报, 2010, 26(6): 1643-1650. LIU F F, DONG X Y, SUN Y. Molecular dynamics simulation of the conformational transition of amyloid peptide 42 inhibited by trehalose[J]. Acta Physico-Chimica Sinica, 2010,26(6): 1643-1650.
[10] NAIKI H, HIGUCHI K, HOSOKAWA M, et al. Fluorometric determination of amyloid fibrils in vitro using the fluorescent dye, thioflavin T1[J]. Analytical Biochemistry, 1989, 177: 244-249. DOI: 10.1016/0003-2697(89)90046-8.
[11] LEVINE H, 3RD. Quantification of beta-sheet amyloid fibril structures with thioflavin T[J]. Methods in Enzymology, 1999, 309: 274-284. DOI: 10.1016/S0076-6879(99)09020-5.
[12] WOLFE L S, CALABRESE M F, NATH A, et al. Protein-induced photophysical changes to the amyloid indicator dye thioflavin T[J]. Proceedings of the National Academy of Sciences of the United States of America, 2010, 107: 16863-16868. DOI: 10.1073/pnas.1002867107.
[13] AMDURSKY N, EREZ Y, HUPPERT D. Molecular rotors: What lies behind the high sensitivity of the thioflavin-T fluorescent marker[J]. Accounts of Chemical Research, 2012, 45: 1548-1557. DOI: 10.1021/ar300053p.
[14] LEVINE H, 3RD. Stopped-flow kinetics reveal multiple phases of thioflavin T binding to alzheimer beta(1-40)amyloid fibrils[J]. Archives of Biochemistry and Biophysics, 1997, 342: 306-316. DOI:10.1006/abbi.1997.0137.
[15] SABATE R, LASCU I, SAUPE S J. On the binding of thioflavin-T to HET-s amyloid fibrils assembled at pH 2[J]. Journal of Structural Biology, 2008, 162: 387-396. DOI:10.1016/j.jsb.2008.02.002.
[16] QIN Z, SUN Y, JIA B H, et al. Kinetic mechanism of thioflavin T binding onto the amyloid fibril of hen egg white lysozyme[J]. Langmuir, 2017, 33: 5398-5405. DOI: 10.1021/acs.langmuir.7b00221.
[17] VERNAGLIA B A, HUANG J, CLARK E D. Guanidine hydrochloride can induce amyloid fibril formation from hen egg-white lysozyme[J]. Biomacromolecules, 2004, 5: 1362-1370. DOI: 10.1021/bm0498979.
[18] SULATSKAYA A I, KUZNETSOVA I M, TUROVEROV K K. Interaction of thioflavin T with amyloid fibrils: Stoichiometry and affinity of dye binding, absorption spectra of bound dye[J]. Journal of Physical Chemistry B, 2011, 115: 11519-11524. DOI: 10.1021/jp207118x.
[19] ZOU Y, LI Y, HAO W, et al. Parallel beta-sheet fibril and antiparallel beta-sheet oligomer: New insights into amyloid formation of hen egg white lysozyme under heat and acidic condition from FTIR spectroscopy[J]. Journal of Physical Chemistry B, 2013, 117: 4003-4013. DOI: 10.1021/jp4003559.
[20] KLUNK W E, JACOB R F, MASON R P. Quantifying amyloid by congo red spectral shift assay[J]. Methods in Enzymology,1999, 309: 285-305. DOI:10.1016/S0076-6879(99)09021-7.
[21] BARTH A, ZSCHERP C. What vibrations tell us about proteins[J]. Quarterly Reviews of Biophysics, 2002, 35: 369-430. DOI: 10.1017/S0033583502003815.

硫代黄素T与溶菌酶淀粉样纤维结合的荧光停流动力学

Fluorescence-detected stopped-flow kinetic investigation of thioflavin T binding onto the amyloid fibril of lysozyme

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价

[1]	叶剑华,于涛,余豪奇,刘会誉,原雪燕. 甲基橙在天然水滑石上的在线吸附周期性规律研究[J]. 河北大学学报(自然科学版), 2023, 43(4): 395-407.
[2]	李妍, 曹思雨,马刚. Hofmeister序列阴阳离子对溶菌酶淀粉样纤维的影响[J]. 河北大学学报(自然科学版), 2023, 43(2): 146-155.
[3]	李雪辰,陈俊宇,贾鹏英,武珈存,冉俊霞. 外加电压参数对大气压等离子体射流形貌的影响[J]. 河北大学学报(自然科学版), 2021, 41(5): 495-502.
[4]	金显春,宋嘉宁. 改性烟曲霉对刚果红的去除动力学、等温线及机制[J]. 河北大学学报(自然科学版), 2019, 39(6): 611-618.
[5]	贾宝环, 于洋,王悦,刘思阳,杜保安,马刚. 盐效应对还原变性溶菌酶聚集的影响[J]. 河北大学学报(自然科学版), 2018, 38(6): 588-595.
[6]	李博,田瑞兰,张炜华 ,刘国欣. 粒子群算法在非线性金融风险模型中的应用[J]. 河北大学学报(自然科学版), 2018, 38(3): 225-231.
[7]	唐然肖,李妍,范胜男,尚洪琳,王春,赵影. 核桃壳粉对阳离子染料结晶紫的吸附特性[J]. 河北大学学报(自然科学版), 2018, 38(3): 254-261.
[8]	刘佳,李娜,郝子岩,姜海勇,李朝晖. 柔性机械手设计与夹取力动态特性仿真分析[J]. 河北大学学报(自然科学版), 2018, 38(2): 119-125.
[9]	代群威,王可,王岩,李刚. 灭活面包酵母菌对水溶液中镉离子的吸附特性[J]. 河北大学学报(自然科学版), 2017, 37(5): 523-530.
[10]	闫佳,冯帆,张永亮,贺亚峰. Oregonator模型中欠扩散斑图动力学[J]. 河北大学学报(自然科学版), 2017, 37(1): 19-23.
[11]	梁诗敏,于涛. 铀在高岭土上的吸附动力学及热力学研究[J]. 河北大学学报(自然科学版), 2016, 36(6): 596-603.
[12]	党伟,刘旭,白晶晶,赵晋津,张连水. CH₃NH₃PbI₃薄膜的光致发光增强效应[J]. 河北大学学报(自然科学版), 2016, 36(5): 462-467.
[13]	李昊一,李冬敏,马刚,商艳丽,孙英. 牛血清白蛋白聚集体的分解动力学[J]. 河北大学学报(自然科学版), 2016, 36(1): 35-42.
[14]	白海鑫,刘小花,王瑾,金秋. 氧化石墨烯对水溶液中中性红的吸附行为[J]. 河北大学学报(自然科学版), 2015, 35(5): 473-479.
[15]	于涛,夏天,殷志成,尚小琦. Eu(Ⅲ)在红壤及其实性成分上的吸附动力学及热力学[J]. 河北大学学报(自然科学版), 2014, 34(6): 616-623.