高等学校化学学报 ›› 2018, Vol. 39 ›› Issue (3): 476-481.doi: 10.7503/cjcu20170316

• 有机化学 • 上一篇    下一篇

神经红蛋白突变体H64V与叠氮离子的作用

高朝辉, 李海丽, 王蕾, 邱章磊, 穆建帅, 李连之()   

  1. 聊城大学化学化工学院, 聊城 252059
  • 收稿日期:2017-05-19 出版日期:2018-03-10 发布日期:2018-01-23
  • 作者简介:联系人简介: 李连之, 男, 博士, 教授, 主要从事生物无机化学研究. E-mail:lilianzhi1963@163.com
  • 基金资助:
    国家自然科学基金(批准号: 20471025, 21142003)和山东省自然科学基金(批准号: ZR2011BL002)资助

Interaction of Neuroglobin Mutant H64V with Azide Anion

GAO Chaohui, LI Haili, WANG Lei, QIU Zhanglei, MU Jianshuai, LI Lianzhi*()   

  1. School of Chemistry and Chemical Engineering, Liaocheng University, Liaocheng 252059, China
  • Received:2017-05-19 Online:2018-03-10 Published:2018-01-23
  • Contact: LI Lianzhi E-mail:lilianzhi1963@163.com
  • Supported by:
    † Supported by the National Natural Science Foundation of China(Nos.20471025,21142003) and the Natural Science Foundation of Shandong Province, China(No.ZR2011BL002)

摘要:

将64位组氨酸(His64)突变为缬氨酸(Val), 构建了神经红蛋白(Ngb)突变体H64V基因并进行了蛋白的表达纯化, 突变使Ngb的血红素Fe由六配位结构变成五配位结构. 利用光谱法研究了突变体H64V与叠氮离子()的相互作用. 紫外-可见吸收光谱表明, H64V与键合后其Soret带吸收峰由406 nm红移至418 nm; 荧光光谱表明可使H64V的荧光发生静态猝灭. 计算得到了H64V与键合的平衡解离常数和热力学参数. 另外, 还研究了不同阴离子对H64V与结合的影响, 发现阴离子的存在减弱了与H64V之间的结合力.

关键词: 神经红蛋白, 突变体H64V, 叠氮离子, 光谱法

Abstract:

Mutating the histidine 64 to valine, the gene of Neuroglobin(Ngb) mutant H64V was constructed and its protein was expressed and purified. This mutation changed the coordination geometry of neuroglobin from hexa- to penta-coordinated. The interaction of neuroglobin mutant H64V with azide anion() was investigated by spectroscopy. UV-Vis absorbance spectra showed that the binding of mutant H64V to induced the change of the Soret-band absorption peak from 406 nm to 418 nm. Fluorescence spectra indicated that the fluorescence of mutant H64V was quenched by through static quenching mechanism. The binding equilibrium dissociation constant and thermodynamic parameters between mutant H64V and were calculated. In addition, the effects of different anions on their binding were studied, the results showed that the anions reduced the binding ability between mutant H64V and azide anion.

Key words: Neuroglobin, Mutant H64V, Azide anion, Spectroscopy

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