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光诱导高铁肌红蛋白还原的光谱法研究
  • 期刊名称:化学学报
  • 时间:0
  • 页码:1559-1564
  • 语言:中文
  • 分类:O644.1[理学—物理化学;理学—化学]
  • 作者机构:[1]大连大学生物工程学院,大连116622, [2]辽宁省生物有机化学重点实验室大连大学,大连116622
  • 相关基金:国家自然科学基金(No.20871024); 辽宁省高校创新团队项目(Nos.2006T002 2008T005 2009T003); 辽宁省教育厅项目(Nos.2009A069 2009A071); 大连市科技计划项目(No.2008E11SF170)资助项目
  • 相关项目:光诱导肌红蛋白的去氧、结合氧及传能机制研究
中文摘要:

采用紫外-可见吸收、荧光及圆二色(CD)等光谱法为主要分析手段,对光诱导高铁肌红蛋白(metmyoglobin,metMb)还原及其引起的蛋白结构变化进行了研究.采用氙灯430 nm的单色光和紫外灯对高铁肌红蛋白溶液进行光照实验,实验结果显示,在metMb的Q吸收带545 nm和580 nm附近吸收峰强度显著增强.通过验证性实验,证明光照后metMb发生了还原反应.验证性实验包括:metMb中加入还原剂连二亚硫酸钠实验;光照后样品加入氧化剂铁氰化钾的可逆性实验;光照的同时通入CO实验.进一步讨论表明,采用氙灯430 nm的单色光诱导metMb还原的机理为光诱导的分子内电子转移.此外,对光还原反应后的样品进行同步荧光和CD光谱分析,发现蛋白质色氨酸残基微环境极性增加,α-螺旋的含量由63%下降到57%,而β-折叠的含量增加了7%.

英文摘要:

The photoreduction of metmyoglobin(metMb) and its effects on the changes of protein structure were investigated by UV-visible absorbance,fluorescence and circular dichroism(CD) spectra.MetMb in Na2HPO4-NaH2PO4 buffer were irradiated with xenon lamp at 430 nm and ultraviolet lamp.The results in-dicated that the absorbance intensity at Q band(545 nm and 580 nm) increased evidently after illumination.Through experimental verification,metMb had been reduced after the illumination.The verifying experi-ment included: adding sodium dithionite to metMb;adding potassium ferricyanide to the sample after the il-lumination;irradiating meanwhile adding CO to the sample.Through further discussion,it can be seen that the photoreduction mechanism of metMb with xenon lamp at 430 nm was photoinduced intramolecular elec-tron transfer.In addition,the results of synchronous fluorescence and CD spectra of the sample after irradia-tion suggested that the polarity of microenvironment of tryptophan residues increased and the secondary structure of protein(CD data) varied with α-helices reduction from 63% to 57%,and β-sheet increase by 7%.

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