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Molecular property and functional control of chaperone-like polymer

类伴侣聚合物的分子特性及功能调控

基本信息

批准号：
16550109
负责人：
TANAKA Naoki
金额：
$ 2.37万
依托单位：
Kyoto Institute of Technology
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2004
资助国家：
日本
起止时间：
2004 至 2005
项目状态：
已结题

项目摘要

The irreversible protein denaturation due to the aggregation is serious problem in the research of protein structure. Molecular chaperones, member of heat shock protein, prevent the protein aggregate in vivo through the weak protein integration. It has been shown that phospholipid polymers (MPC polymer) also weakly interact proteins, and we used these polymers as the artificial molecular chaperone. We have design several MPC copolymers, and screened by there chaperone-like activity. The activity for the refolding of β-galactosidase, malate dehydrogenase, MHC Class II, luciferase GFP and rhodanese have been examined. The copolymer with butylmethacrylate (PMB polymer) showed high performance for the refolding of these proteins.The in vitro protein synthesis is one of the effective methods for this purpose, but this method has weak point of the low yield. To construct a highly efficient in vitro protein synthesis system, the effects PMB polymer were examined. Although PMB polymer showed chaperone-like activity, the yield of rhodanese in the in vitro protein synthesis was not enhanced by this polymers. Another new design of the MPC polymer is necessary for the efficient in vitro protein synthesis system. Furthermore, we examined the effect of MPC copolymers on the amyloid fibril formation of insulin. We found that hydrophilic copolymers accelerated the amyloid formation rate, and the hydrophobic copolymers reduced the total amount of amyloid fibril. But so far, MPC copolymers don't have sufficient ability to control amyloid fibril structure. We will improve the functional capability of MPC copolymer by the arrangement of copolymerization ratio and molecular weight.

蛋白质聚集引起的不可逆变性是蛋白质结构研究中的一个重要问题。分子伴侣是热休克蛋白的成员，通过弱的蛋白质整合阻止蛋白质在体内聚集。研究表明磷脂聚合物（MPC聚合物）也与蛋白质弱相互作用，我们将这些聚合物用作人工分子伴侣。我们设计了几种MPC共聚物，并对其进行了分子伴侣活性筛选。已经检查了β-半乳糖苷酶、苹果酸脱氢酶、MHC II类、荧光素酶GFP和rhodanese的重折叠活性。与甲基丙烯酸丁酯的共聚物（PMB聚合物）对这些蛋白质的复性表现出很好的性能，体外蛋白质合成是实现这一目的的有效方法之一，但该方法存在收率低的缺点。为了构建一个高效的体外蛋白质合成系统，研究了PMB聚合物的作用。虽然PMB聚合物表现出分子伴侣样活性，但在体外蛋白质合成中罗丹酸的产率并没有被该聚合物提高。MPC聚合物的另一种新设计对于有效的体外蛋白质合成系统是必要的。此外，我们研究了MPC共聚物对胰岛素淀粉样纤维形成的影响。我们发现，亲水性共聚物加速了淀粉样蛋白的形成速率，而疏水性共聚物减少了淀粉样蛋白原纤维的总量。但到目前为止，MPC共聚物还没有足够的能力来控制淀粉样纤维的结构。通过对MPC共聚物的共聚比和分子量的调整，可以提高MPC共聚物的功能性能。