PDZ、SH3结构域结合特性的研究

发布时间：2018-01-16 02:14

  本文关键词：PDZ、SH3结构域结合特性的研究　出处：《中国协和医科大学》2009年硕士论文　论文类型：学位论文

  更多相关文章： 蛋白质相互作用 结构域 PDZ结构域 SH3结构域 结合特性 酵母双杂交 随机多肽文库 验证筛选 PDLIM2蛋白 PDZK2蛋白 FYN蛋白 非脯氨酸文库

【摘要】： 蛋白质在细胞的生命活动中扮演着重要角色,但是,其功能的发挥却并非依靠单个蛋白质独立的作用。它们在细胞中通常与其他蛋白质相互作用形成大的复合体,在特定的时间和空间内完成特定的功能,蛋白质之间的相互作用才是蛋白质执行其功能的途径。 本论文主要研究了介导蛋白质相互作用网络的PDZ结构域和SH3结构域的结合特性。 本文研究了PDLIM2蛋白中的PDZ结构域的结合特性。利用酵母双杂交技术筛选随机多肽文库,我们验证了PDLIM2-PDZ可识别x-x-W-L配体；此外还发现了K-H-K-V配体,是对PDLIM2-PDZ蛋白识别规律的补充。 论文第二部分研究了PDZK2蛋白中PDZ1的结合特性。根据PDZK2 PDZ1结构域与PDZK1 PDZ1同源的性质,我们直接利用酵母双杂交技术筛选构建的PDZ配体库来研究PDZK2 PDZ1的结合特性。验证筛选方法较常规文库筛选具有高效性和针对性,可以快速研究相关结构域的结合特性。验证筛选文库相对于其他文库筛选方法可以得到除阳性配体外的阴性序列,并且阴性序列和阳性配体对比总结可以得到更全面的结合特性。 论文第三部分成功构建新型非脯氨酸的随机多肽文库。我们尝试利用新型文库研究结合非脯氨酸的SH3结构域的结合特性,发现了FYN SH3结构域的一个非脯氨酸结合序列VSLVKLFF。
[Abstract]:Proteins play an important role in cell life, but. Their function is not independent of a single protein. They usually interact with other proteins in cells to form large complexes that perform specific functions in specific time and space. The interaction between proteins is the way for proteins to perform their functions. In this paper, the binding properties of PDZ domain and SH3 domain mediated protein interaction network are studied. The binding characteristics of PDZ domain in PDLIM2 protein were studied and the random polypeptide library was screened by yeast two-hybrid technique. We have verified that PDLIM2-PDZ can recognize x-x-W-L ligand. In addition, K-H-K-V ligand was found, which is a complement to the recognition rule of PDLIM2-PDZ protein. In the second part, the binding properties of PDZ1 in PDZK2 protein were studied. According to the homology between PDZK2 PDZ1 domain and PDZK1 PDZ1. We directly used yeast two-hybrid technology to screen the constructed PDZ ligands to study the binding characteristics of PDZK2 PDZ1. The binding characteristics of related domains can be studied quickly. Compared with other screening methods, negative sequences in vitro can be obtained by verifying screening libraries. And the comparison of negative sequence and positive ligand can obtain more comprehensive binding characteristics. In the third part of the thesis, we successfully construct a novel non-proline random polypeptide library. We try to use the new library to study the binding characteristics of the SH3 domain combined with non-proline. A non-proline binding sequence of FYN SH3 domain VSLVKLFF was found.
【学位授予单位】：中国协和医科大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R341

【参考文献】

中国期刊全文数据库 前1条

1 ;FINDING POTENTIAL LIGANDS FOR PDZ DOMAINS BY TAILFIT, A JAVA PROGRAM[J];Chinese Medical Sciences Journal;2004年02期

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本文编号：1431088