D60K Mb定点突变对肌红蛋白结构及荧光量子产率的影响

doi:10.7517/j.issn.1674-0475.2017.05.001

影像科学与光化学 ›› 2017, Vol. 35 ›› Issue (6): 788-796.DOI: 10.7517/j.issn.1674-0475.2017.05.001

D60K Mb定点突变对肌红蛋白结构及荧光量子产率的影响

唐乾^1,2, 李如玉^2,3, 职秋雁^1,2, 曹洪玉^1,2, 王立皓^2,3, 郑学仿^1,2

1. 大连大学生命科学与技术学院, 辽宁大连 116622;
2. 大连大学辽宁省生物有机化学重点实验室, 辽宁大连 116622;
3. 大连大学环境与化学工程学院, 辽宁大连 116622

收稿日期:2016-12-08 修回日期:2017-03-06 出版日期:2017-11-15 发布日期:2017-11-15
通讯作者: 郑学仿, 唐乾
基金资助:
国家自然科学基金项目（21571025，21271036，21601024）资助

Research on the Effect of Asp60Lys on the Structure and Fluorescence Quantum Yield of Myoglobin

TANG Qian^1,2, LI Ruyu^2,3, ZHI Qiuyan^1,2, CAO Hongyu^1,2, WANG Lihao^2,3, ZHENG Xuefang^1,2

1. College of Life Science and Biotechnology, Dalian University, Dalian 116622, Liaoning, P. R. China;
2. Liaoning Key Laboratory of Bio-organic Chemistry, Dalian University, Dalian 116622, Liaoning, P. R. China;
3. School of Environmental and Chemical Engineering, Dalian University, Dalian 116622, Liaoning, P. R. China

Received:2016-12-08 Revised:2017-03-06 Online:2017-11-15 Published:2017-11-15

摘要/Abstract

摘要：

为了探究肌红蛋白表面60位的Asp突变为Lys后对蛋白荧光强度的影响，在模拟生理条件下，利用荧光法和圆二色法对WT Mb及D60K Mb的结构及荧光光谱特性进行了对照研究，结果表明D60K Mb的荧光量子产率远大于WT Mb。通过研究pH值和结构对荧光量子产率的影响，发现Mb 60位天冬氨酸（Asp）突变为赖氨酸（Lys）后，蛋白表面电荷发生改变，导致蛋白的荧光量子产率明显增强；同时使蛋白内部的芳香族氨基酸较原来更暴露，极性增强，激发态分子的去激发过程由原来的无辐射能量转移为主转变为辐射能量转移为主，最终导致荧光强度增强。

关键词: 肌红蛋白, D60K Mb, 光谱, 结构, 荧光量子产率

Abstract:

To study the role of surface-charged Asp60 in the fluorescence intensity, under simulative physiological conditions of animal body, the difference in structure and fluorescence spectral characteristics between Mb(WT) and Mb(D60K) was studied by fluorescence spectroscopy and circular dichroism. The results indicated that the fluorescence quantum yield of (D60K) was far more than that of Mb, which may correlate with pH and the structure of Mb. With a further research, this phenomenon was caused by the change of external charge when amino acid Asp60 of myoglobin was replaced by Lys, which enhanced the polarity and exposure extent of aromatic amino acids of myoglobin, and the decay process of myoglobin molecules which were excited from non-radiative decay to radiative decay.

Key words: myoglobin, mutant(D60k), spectroscopy, structure, fluorescence quantum yield

唐乾, 李如玉, 职秋雁, 曹洪玉, 王立皓, 郑学仿. D60K Mb定点突变对肌红蛋白结构及荧光量子产率的影响[J]. 影像科学与光化学, 2017, 35(6): 788-796.

TANG Qian, LI Ruyu, ZHI Qiuyan, CAO Hongyu, WANG Lihao, ZHENG Xuefang. Research on the Effect of Asp60Lys on the Structure and Fluorescence Quantum Yield of Myoglobin[J]. Imaging Science and Photochemistry, 2017, 35(6): 788-796.

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D60K Mb定点突变对肌红蛋白结构及荧光量子产率的影响

Research on the Effect of Asp60Lys on the Structure and Fluorescence Quantum Yield of Myoglobin

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