黄牛肝菌凝集素的化学修饰与荧光光谱研究
Chemical modification and fluorescence analysis of lectin from Boletus fulvus
作者：王楠(四川大学生命科学学院生物资源与生态环境教育部重点实验室)；祁伟(四川大学生命科学学院生物资源与生态环境教育部重点实验室)；周立(四川大学生命科学学院生物资源与生态环境教育部重点实验室)；鲍锦库(四川大学生命科学学院生物资源与生态环境教育部重点实验室)
Author：WANG Nan(Key laboratory of Bio resources and Eco environment of Ministry of Education,College of Life Sciences, Sichuan University)；QI Wei(Key laboratory of Bio resources and Eco environment of Ministry of Education,College of Life Sciences, Sichuan University)；ZHOU Li(Key laboratory of Bio resources and Eco environment of Ministry of Education,College of Life Sciences, Sichuan University)；BAO Jin-Ku(Key laboratory of Bio resources and Eco environment of Ministry of Education,College of Life Sciences, Sichuan University)
收稿日期：2013-12-26 年卷（期）页码：2015,52(3):657-662
期刊名称：四川大学学报: 自然科学版
Journal Name：Journal of Sichuan University (Natural Science Edition)
关键字：黄牛肝菌凝集素; 化学修饰; 荧光光谱; 凝血活性; 构象
Key words：Boletus fulvus lectin; Chemical modification; Fluorescence spectrum; Hemagglutination activity; Conformation
基金项目：国家自然科学基金(81173093, 30970643, J1103518)
中文摘要
对黄牛肝菌凝集素(Boletus fulvus lectin, BFL)进行氨基酸化学修饰, 结果表明, Trp残基、Tyr残基及Ser/Thr残基的修饰都会对BFL的凝血活性产生影响, 表明这些残基是BFL凝血活性所必需的, 可能参与了其凝血活性中心的构成. 而Arg残基修饰后, BFL的凝血活性未发生改变, 表明Arg不是维持BFL凝血活性的必需基团. 内源荧光光谱分析结果表明, BFL所发射的荧光是由Trp残基贡献的, 且Trp残基在BFL分子中所处的微环境极性较弱, 屏蔽于一定的构象当中. 温度、pH值及变性剂对BFL内源荧光光谱的影响与基本理化性质的实验结果一致. 氨基酸修饰对荧光光谱的影响表明, Trp残基修饰会引起BFL內源荧光光谱较大变动, 而Tyr、Ser/Thr残基修饰对BFL內源荧光光谱影响不大.
英文摘要
Chemical modification of amino acid residues of Boletus fulvus lectin (BFL) showed that treating purified BFL with amino acid modifying reagents specific for Trp, Tyr and Ser/Thr could affect the hemagglutination activity of BFL. These results indicated that these residues played important roles in the hemagglutination activity of BFL, and might be essential for constituting the active center of BFL, whereas Arg might not be essential for constituting the active center of BFL. Moreover, the fluorescence spectrum of native BFL suggested that Trp residues made great contribution to the fluorescence of BFL, and they might locate in the hydrophobic microenvironment of BFL and be buried in the hydrophobic core of the lectin. The influences of temperature, pH and denaturants on the fluorescence spectrum of BFL were consistent with the hemagglutination assays. The experiment of amino acid modification suggested that modification of Trp significantly affected the fluorescence spectrum of BFL, whereas modification of Tyr and Ser/Thr induced little change on the fluorescence spectrum of BFL.
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