物理化学学报 >> 2003, Vol. 19 >> Issue (09): 834-838.doi: 10.3866/PKU.WHXB20030911

研究论文 上一篇    下一篇

色氨酸标记的GCN4单体肽与DNA位点的分子识别

王旭;邓巍;曹傲能;来鲁华   

  1. 分子动态与稳态结构国家重点实验室, 北京大学化学与分子工程学院, 北京　100871
  • 收稿日期:2003-03-24 修回日期:2003-05-06 发布日期:2003-09-15
  • 通讯作者: 来鲁华 E-mail:lhlai@pku.edu.cn

Molecular Recognition of GCN4 Monomeric Peptide Tryptophane Mutant with Its DNA Target Sites

Wang Xu;Deng Wei;Cao Ao-Neng;Lai Lu-Hua   

  1. State Key Laboratory for Structural Studies of Stable and Unstable Species, College of Chemistry and Molecular Engineering, Peking University, Beijing　100871
  • Received:2003-03-24 Revised:2003-05-06 Published:2003-09-15
  • Contact: Lai Lu-Hua E-mail:lhlai@pku.edu.cn

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摘要： 酵母转录激活因子GCN4调控细胞中氨基酸的生物合成, 是典型的含bZIP结构域的DNA结合蛋白.本文合成了天然蛋白GCN4的碱性区（226-252）, 并在其N末端引入色氨酸残基W, 做为单体肽GCN4-W.圆二色(CD)实验表明, 突变后的单体肽仍能序列特异性识别DNA结合位点AP-1和ATF/CREB.用荧光滴定方法获得了GCN4-W与DNA位点结合形成复合物的表观解离常数.

关键词: 酵母转录激活因子(GCN4), 圆二色(CD), 荧光滴定, 解离常数

Abstract: The transcriptional activator GCN4 plays a key role in the regulation of amino acid anabolism in yeast. It is a DNA binding protein that has typical bZIP domain. We synthesized the basic region (226-252) of natural protein GCN4, and introduced a Trp residue at the N-terminal. CD experimental data show that the synthesized monomer peptide, GCN4-W, can still specifically recognize the DNA target sites AP-1 and ATF/CREB. We also obtained the apparent dissociation constant of the peptide-DNA complex of this monomer recognizing DNA site by fluorescence titration method.

Key words: Transcriptional activator(GCN4), CD, Fluorescence titration, 　Dissociation constant