本文關(guān)鍵詞：PDZ、SH3結(jié)構(gòu)域結(jié)合特性的研究　出處：《中國協(xié)和醫(yī)科大學(xué)》2009年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 蛋白質(zhì)相互作用 結(jié)構(gòu)域 PDZ結(jié)構(gòu)域 SH3結(jié)構(gòu)域 結(jié)合特性 酵母雙雜交 隨機多肽文庫 驗證篩選 PDLIM2蛋白 PDZK2蛋白 FYN蛋白 非脯氨酸文庫

【摘要】： 蛋白質(zhì)在細(xì)胞的生命活動中扮演著重要角色,但是,其功能的發(fā)揮卻并非依靠單個蛋白質(zhì)獨立的作用。它們在細(xì)胞中通常與其他蛋白質(zhì)相互作用形成大的復(fù)合體,在特定的時間和空間內(nèi)完成特定的功能,蛋白質(zhì)之間的相互作用才是蛋白質(zhì)執(zhí)行其功能的途徑。 本論文主要研究了介導(dǎo)蛋白質(zhì)相互作用網(wǎng)絡(luò)的PDZ結(jié)構(gòu)域和SH3結(jié)構(gòu)域的結(jié)合特性。 本文研究了PDLIM2蛋白中的PDZ結(jié)構(gòu)域的結(jié)合特性。利用酵母雙雜交技術(shù)篩選隨機多肽文庫,我們驗證了PDLIM2-PDZ可識別x-x-W-L配體；此外還發(fā)現(xiàn)了K-H-K-V配體,是對PDLIM2-PDZ蛋白識別規(guī)律的補充。 論文第二部分研究了PDZK2蛋白中PDZ1的結(jié)合特性。根據(jù)PDZK2 PDZ1結(jié)構(gòu)域與PDZK1 PDZ1同源的性質(zhì),我們直接利用酵母雙雜交技術(shù)篩選構(gòu)建的PDZ配體庫來研究PDZK2 PDZ1的結(jié)合特性。驗證篩選方法較常規(guī)文庫篩選具有高效性和針對性,可以快速研究相關(guān)結(jié)構(gòu)域的結(jié)合特性。驗證篩選文庫相對于其他文庫篩選方法可以得到除陽性配體外的陰性序列,并且陰性序列和陽性配體對比總結(jié)可以得到更全面的結(jié)合特性。 論文第三部分成功構(gòu)建新型非脯氨酸的隨機多肽文庫。我們嘗試?yán)眯滦臀膸煅芯拷Y(jié)合非脯氨酸的SH3結(jié)構(gòu)域的結(jié)合特性,發(fā)現(xiàn)了FYN SH3結(jié)構(gòu)域的一個非脯氨酸結(jié)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.
【學(xué)位授予單位】：中國協(xié)和醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2009
【分類號】：R341

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本文編號：1431088