Alpha Crystallin Chaperone Function in the Zebrafish

描述(申请人提供)：脊椎动物眼睛的30% 晶状体是由小分子热休克蛋白α-晶状体蛋白组成。这种蛋白质 起到分子伴侣的作用，防止应激诱导的聚集 可导致白内障的变性蛋白质。这一点的中心前提是 认为α-晶状体蛋白的伴侣活性依赖于 温度，它的氨基酸组成已经进化到保持 伴侣在不同的热环境中发挥作用。α-晶状体蛋白由 由基因复制事件产生的两个亚基，αA和αB 并具有不同的伴侣属性。这项提案旨在确定 αA-和α-蛋白的结构与伴侣功能的关系 斑马鱼中的α-晶状体蛋白。这项研究将首次进行化验 时间，来自外温脊椎动物的α-晶体蛋白的伴侣功能。 这一信息将直接与结构和伴侣进行比较 人αA和αB-晶体蛋白的性质决定伴侣蛋白是否存在 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 在小的热休克蛋白的功能和白内障，并将建立基本 热适应的进一步研究需要的知识 α-晶状体蛋白。 提出的目标将使用跨学科的方法来实现。 结合了分子生物学、蛋白质生物化学、生态学和进化论。这个 高级调查员在鱼类进化、鱼类生态和 分子技术使他成为唯一有资格进行这项工作的人 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.