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小肽对Hoechst 33258与DNA特异部位结合的竞争性作用

杨铭, 朱树梅, 黄艳萍, 胡齐悦, 王夔   

  1. 北京医科大学 天然药物及仿生药物国家重点实验室, 北京 100083
  • 收稿日期:1996-03-27 修回日期:1996-06-24 出版日期:1996-09-15 发布日期:1996-09-15

Studies of Site Specific DNA Binding of Small Peptides by Competitive Assays with Hoechst 33258

Ming Yang, Shu-Mei Zhu, Yan-Ping Huang, Qi-Yue Hu, Kui Wang   

  1. National Research Laboratories of Natural and Biomimetic Drugs; Beijing Medical University; Beijing 100083
  • Received:1996-03-27 Revised:1996-06-24 Online:1996-09-15 Published:1996-09-15

摘要: 为了了解小肽如何识别DNA特异的结合位点, 我们研究了两个小肽YRGGHDNAHoechst 33258特异结合的竞争性抑制作用。对Lineweaver-Burk图的分析显示当Hoechst 33258的浓度在3.66 (10-9 mol/L to 1.09 ( 10-8 mol/L范围内, 这种作用是典型的竞争性抑制作用, YR结合在DNA小沟区(AT富集区), 结合常数为1.02 ( 108 (mol/L)-1, GGHAT富集区结合特异性在浓度高时明显减弱, 因为它也能与GC碱基结合。

关键词: DNA, 小肽, 竞争性抑制剂, 部位特异性结合

Abstract: With a view to finding out precisely how small peptides recognize a particular binding site of DNA, we have accomplished DNA binding studies of two peptides, H-Tyr-Arg-OH (YR) and H-Gly-Gly-His-OH (GGH) by using measurements in comparison with the binding between DNA and Hoechst 33258. The inhibition mode by YR and GGH to DNA binding of Hoechst 33258 was analyzed by Lineweaver-Burk plot which shows the plot of typical competitive inhibition at concentration of Hoechst 33258 from 3.66 (10-9 mol/L to 1.09 ( 10-8 mol/L. And it is concluded that YR binds to DNA in its minor groove (AT rich regions) with a binding constant K = 1.02 ( 108 (mol/L)-1. The GGH (s specificity is reduced at high concentration because it can also bind GC base pair.

Key words: DNA, Peptide, Competitive inhibitor, Site specific binding

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