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化学学报
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化学学报 ›› 2006, Vol. 64 ›› Issue (13): 1349-1354. 上一篇    下一篇

研究论文

柚皮素、柚皮苷与溶菌酶相互作用的荧光光谱法研究

杨冉1，屈凌波*,2，陈晓岚2，李建军2，李萍*,1   

  1. (1中国药科大学生药教研室 现代中药教育部重点实验室 南京 210038)
    (2郑州大学化学系 化学生物重点实验室 郑州 450052)
  • 投稿日期:2005-07-22 修回日期:2006-03-27 发布日期:2006-07-14
  • 通讯作者: 屈凌波

Studies on the Interaction of Lysozyme with Naringein and Naringin by Fluorescence Spectroscopy

YANG Ran1, QU Ling-Bo*,2, CHEN Xiao-Lan2, LI Jian-Jun2, LI Ping*,1   

  1. (1 Key Laboratory of Modern Chinese Medicines, Ministry of Education, Department of Pharmacognosy,
    China Pharmaceutical University, Nanjing 210038)
    (2 Key Laboratory of Chemical Biology, Department of Chemistry, Zhengzhou University, Zhengzhou 450052)
  • Received:2005-07-22 Revised:2006-03-27 Published:2006-07-14
  • Contact: QU Ling-Bo

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1512

被引次数

65

应用荧光光谱法研究了50%甲醇/水体系中柚皮素、柚皮苷与溶菌酶分子间的结合反应. 以Lineweaver-Burk双倒数方程和能量传递原理分别计算了两者与溶菌酶反应的结合常数(K)和结合距离(r): K柚皮素25 ℃＝4.00×104, K柚皮苷25 ℃＝3.48×104; r柚皮素＝3.21 nm, r柚皮苷＝3.30 nm, 以及由热力学参数的计算判断了两种分子与溶菌酶之间的作用力类型. 结果表明: 柚皮素、柚皮苷均能与溶菌酶以疏水作用相结合形成非共价化合物, 从而导致溶菌酶内在荧光的静态猝灭; 相对柚皮素, 柚皮苷与溶菌酶的结合距离增大, 作用强度减弱, 表明黄酮分子上多糖的取代不利于黄酮分子与蛋白之间的亲和作用. 根据Haslam等提出的多酚-蛋白质反应模型, 从分子水平初步探讨了糖取代对黄酮分子与蛋白相互作用减弱的原因.

关键词: 荧光光谱法, 溶菌酶, 柚皮素, 柚皮苷, 能量传递

The interactions between lysozyme (Lys) and naringein or naringin were studied by fluorescence spectroscopy under 50% CH3OH/water circumstance. The binding constants and bind distance of naringein- Lys and naringin-Lys were calculated according to Lineweaver-Burk equation and Föster’ energy transfer theory, which were shown as KNaringein25 ℃＝4.00×104, rNaringein＝3.21 nm and KNaringin25 ℃＝3.48×104, rNaringin＝3.30 nm, respectively. The results showed that both of them could form non-covalent compounds with Lys mainly through hydrophobic interaction force. Relative to naringein, the binding affinity of naringin to Lys decreased obviously. These suggested that the sugar substitution in the structure of flavonoid is not helpful to the binding of flavonoid-protein. According to the polyphen-protein interaction mode presented by Haslam, the reason resulted in weaker bonding affinity of naringin to Lys was analysed in the level of molecule.

Key words: fluorescence spectroscopy, lysozyme, naringein, naringin, energy transfer