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.

由于聚集而导致的不可逆蛋白质变性是蛋白质结构研究中的严重问题。热休克蛋白的成员分子伴侣通过弱蛋白整合来防止体内蛋白质聚集体。已经表明，磷脂聚合物（MPC聚合物）也弱相互作用的蛋白质，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，并通过伴侣样活动进行筛选。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。与丁基丙烯酸酯（PMB聚合物）的共聚物对这些蛋白质的再折叠表现出很高的性能。体外蛋白质合成是用于此目的的有效方法之一，但是该方法的产量较低。为了构建高效的体外蛋白质合成系统，检查了PMB聚合物的效果。尽管PMB聚合物表现出类似伴侣的活性，但该聚合物并未增强体外蛋白质合成中的罗达烷。 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需的。此外，我们研究了MPC共聚物对胰岛素淀粉样蛋白纤维形成的影响。我们发现亲水共聚物加速了淀粉样蛋白的形成速率，疏水共聚物降低了淀粉样蛋白原纤维的总量。但是到目前为止，MPC共聚物没有足够的控制淀粉样蛋白原纤维结构的能力。我们将通过共聚比和分子量的排列来提高MPC共聚物的功能能力。