基础医学与临床 ›› 2009, Vol. 29 ›› Issue (1): 14-18.

• 研究论文 • 上一篇    下一篇

通过筛选随机多肽文库研究Veli3 PDZ结构域配体结合特性

田瑞 马素参 杨涛 高友鹤   

  1. 中国医学科学院基础医学研究所 中国医学科学院基础医学研究所 中国医学科学院基础医学研究所 中国医学科学院基础医学研究所生理和病理生理系
  • 收稿日期:2008-05-19 修回日期:2008-06-03 出版日期:2009-01-25 发布日期:2009-01-25
  • 通讯作者: 田瑞

Characterization of ligand-binding properties of PDZ domain of Veli3 by screening random peptide library

Rui TIAN, Su-can MA, Tao YANG, You-he GAO   

  1. Institute of Basic Medical Science, CAMS &School of Basic Medicine, PUMC Institute of Basic Medical Science, CAMS &School of Basic Medicine, PUMC Institute of Basic Medical Science, CAMS &School of Basic Medicine, PUMC Institute of Basic Medical Sciences,CAMS & PUMC
  • Received:2008-05-19 Revised:2008-06-03 Online:2009-01-25 Published:2009-01-25
  • Contact: Rui TIAN,

摘要: 目的 研究Veli3 PDZ结构域配体的结合特性。方法 用Veil3 PDZ为诱饵蛋白,用酵母双杂交方法筛选随机多肽文库。结合生物信息学方法在各种数据库中检索与Veli3 PDZ识别规律相符合的天然人类蛋白质。然后根据蛋白质的功能和细胞定位等性质选择14个潜在新配体用酵母双杂交验证相互作用。文献检索与Veli3 PDZ配体结合的其他PDZ蛋白。结果 Veli3 PDZ结构域配体C-末端保守的氨基酸序列通式可以表示为:[E/X][S/T]X[V/I/L]-COOH(X表示任意氨基酸)。这是ClassⅠPDZ结构域配体的特点。用酵母双杂交实验证实14个生物信息方法预测的潜在配体中有6个配体与Veli3相互作用。另一个PDZ蛋白PSD-95与已报道的Veli3 PDZ配体和本研究新发现的潜在配体有多个相互作用。 结论 新发现的6个Veli3PDZ潜在配体为进一步研究Veli3 的生物学功能提供新的线索。另外Veli3与PSD-95可能易于同时竞争结合同一配体,其生物学意义还有待进一步研究。

关键词: 蛋白质相互作用, PDZ结构域, 酵母双杂交, 生物信息学

Abstract: Objective To investigate the ligand-binging characteristics of Veli3 PDZ. Methods Random peptide library was screened using yeast two-hybrid method with Veli3 PDZ as bait. In combination with bioinformatics method all the potential ligands in human proteome have been predicted by searching human databases with the consensus-binding sequences. Fourteen native human proteins predicted as ligands were chosen by their cellular locations and biological functions for validating protein interaction in yeast two-hybrid system. Results The C-terminal consensus sequences for the Veil3 PDZ binding is [E/X][S/T]X[V/I/L]-COOH (X denotes any amino acid), which indicates that Veli3 PDZ belongs to classⅠ. Six of fourteen native human proteins predicted as ligands were confirmed positive in the yeast two-hybrid system. There are a lot of interactions between PSD-95, another PDZ protein, and the ligands of Veli3 PDZ reported previously and discovered in this study. Conclusion The six novel potential ligands of Veli3 found in this study provide significant clues for discovering biological functions of Veil3 proteins. Moreover, Veli3 PDZ and PSD-95 PDZ may compete with each other to bind the same ligands.

Key words: protein interaction, PDZ domain, yeast two hybrid, bioinformatics