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化学学报
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化学学报 ›› 2008, Vol. 66 ›› Issue (9): 1042-1046. 上一篇    下一篇

研究论文

聚乙烯醇与溶菌酶的相互作用及其对溶菌酶构象的影响

王改珍2，贺进田*,1，闫慧源1，周志涛1，周香莲1，侯笑娜1，崔艳2   

  1. (1河北师范大学生命科学院 石家庄 050016)
    (2河北科技大学环境科学与工程学院 石家庄 050018)
  • 投稿日期:2007-09-26 修回日期:2007-11-19 发布日期:2008-05-14
  • 通讯作者: 贺进田

Interaction between PVA and Lysozyme and Its Influence on the Conformation of Lysozyme

WANG Gai-Zhen2 HE Jin-Tian*1 YAN Hui-Yuan1 ZHOU Zhi-Tao1 ZHOU Xiang-Lian1 HOU Xiao-Na1 CUI Yan2   

  1. (1 College of Life Science, Hebei Normal University, Shijiazhuang 050016)
    (2 College of Environmental Science and Engineering, Hebei University of Science and Technology, Shijiazhuang 050018)
  • Received:2007-09-26 Revised:2007-11-19 Published:2008-05-14
  • Contact: HE Jin-Tian

PDF

836

被引次数

20

在生理条件下, 使用凝胶过滤色谱、荧光光谱法、差示扫描量热分析和傅里叶变换红外光谱法(FT-IR)研究了溶菌酶与聚乙烯醇(PVA)的相互作用. 结果表明PVA与溶菌酶结合形成复合物, 在它们的相互作用过程中, 溶菌酶酪氨酸的发射荧光部分被猝灭, 但是, 相互作用并没有改变酪氨酸的微环境; 差示扫描量热分析结果表明, 溶菌酶与PVA之间的相互作用没有破坏溶菌酶的高级结构; 进一步使用红外光谱法结合可增强分辨率的傅里叶去卷积技术和高斯曲线拟合技术共同用于对溶菌酶与PVA复合物冻干粉中溶菌酶酰胺I带的定量分析, 发现冻干粉溶菌酶分子中与分子间相互作用相关的β-折叠组分含量减少了, 但是, 用于衡量冻干状态蛋白质结构完整性的α-螺旋组分含量没有降低. 活性分析结果进一步确认, PVA与溶菌酶的相互作用没有破坏溶菌酶的三级结构.

关键词: 聚乙烯醇, 溶菌酶, 傅里叶变换红外光谱, 凝胶过滤, 荧光光谱

Gel filtration chromatography, fluorescence spectrophotometry, differential scanning calorimetry (DSC) and Fourier transform infrared spectrophotometry (FT-IR) were used to investigate the interaction between polyvinyl alcohol (PVA) and lysozyme under physiological conditions. The results showed that lysozyme formed complex with PVA. During lysozyme interaction with PVA, emission fluorescence derived from tryptophan of lysozyme was partially quenched upon binding to PVA. However, microenvironment of tryptophan was not changed during the binding processs. The results of DSC showed that the interaction of PVA with lysozyme did not destroy advanced structures of lysozyme. Fourier transform infrared spectrometry was combined with resolution enhancement technique Fourier deconvolution and Gaussian curve-fitting procedures to quantitate the spectral information from the amide I bands of lysozyme within the freeze-dried mixture of PVA and lysozyme. The results showed that the interaction of PVA with lysozyme affected only the β-sheet content rather than α-helix , which was usually used as an indicator of the protein structural integrity in lyophilized state. Analysis of activity of the lysozyme showed that interaction of PVA with the lysozyme did not destroy the tertiary structure of the lysozyme.

Key words: polyvinyl alcohol, lysozyme, Fourier transform infrared spectrometry, gel chromatography, fluorescence spectrometry