Alpha Crystallin Chaperone Function in the Zebrafish

描述（由申请人提供）： 脊椎动物眼睛的 30% 晶状体由小的热休克蛋白α-晶状体蛋白组成。这种蛋白质 作为分子伴侣，防止应激诱导的聚集 变性蛋白质可导致白内障。这句话的中心前提是 建议是，α-晶状体蛋白的伴侣活性取决于 温度，并且其氨基酸组成已经进化以维持 不同热环境下的伴侣功能。 α-晶状体蛋白组成 由基因复制事件产生的两个亚基 alphaA 和 alphaB 并具有不同的伴侣特性。该提案旨在确定 αA-和αA-的结构与伴侣功能之间的关系 来自斑马鱼的 αB-晶状体蛋白。这项研究将首次测定 时间，来自变温脊椎动物的α-晶状体蛋白的伴侣功能。 该信息将直接与结构和伴侣进行比较 人 αA 和 αB-晶状体蛋白的特性以确定伴侣是否 功能适应不同物种的生理温度。这些 数据将为大量感兴趣的研究人员提供参考点 在小热休克蛋白功能和白内障中，并将构建基本 进一步研究热适应所需的知识 α-晶状体蛋白。 拟议的目标将通过跨学科方法来实现 结合分子生物学、蛋白质生物化学、生态学和进化论。的 高级研究员在鱼类进化、鱼类生态学和 分子技术使他有资格进行这项工作 跨学科研究项目。

DESCRIPTION (provided by applicant): Up to 30 percent of the vertebrate eye lens is composed of the small heat shock protein alpha-crystallin. This protein acts as a molecular chaperone, preventing the stress-induced aggregation of denatured proteins that can lead to cataracts. The central premise of this proposal is that the chaperone activity of alpha-crystallin is dependent on temperature, and that its amino acid composition has evolved to maintain chaperone function in different thermal environments. Alpha-crystallin consists of two subunits, alphaA and alphaB, that arose from a gene duplication event and have different chaperone properties. This proposal seeks to determine the relationship between the structure and chaperone function of alphaA- and alphaB-crystallin from the zebrafish. This study will assay, for the first time, chaperone function of alpha-crystallin from an ectothermic vertebrate. This information will be directly compared to the structural and chaperone properties of human alphaAand alphaB-crystallin to determine whether chaperone function adapts to the physiological temperatures of different species. These data will provide a reference point for a large body of researchers interested in small heat shock protein function and cataracts, and will build the basic knowledge necessary for further studies on the thermal adaptation of alpha-crystallin. The aims proposed will be accomplished using an interdisciplinary approach combining molecular biology, protein biochemistry, ecology and evolution. The senior investigator's diverse background in fish evolution, fish ecology, and molecular techniques makes him uniquely qualified to carry out this interdisciplinary research project.