MOLECULAR BIOLOGY, STRUCTURES AND FUNCTION OF CRYSTALLINS

晶状体蛋白的分子生物学、结构和功能

• 批准号: 6162363
• 负责人: Graeme J Wistow
• 金额: --
• 依托单位: NATIONAL EYE INSTITUTE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6162363
• 关键词: cataract; cell differentiation; crystallins; cytotoxic T lymphocyte; developmental genetics; gene expression; gene targeting; human genetic material tag; laboratory mouse; lens; lens proteins; linkage mapping; migration inhibition factor; molecular cloning; peptide chemical synthesis; protein structure function; retina; transcription factor

The normal lens depends on abundant proteins known as crystallins for its clarity. Crystallins are multifunctional proteins, recruited to the lens but often retaining other functions important in non-lens tissues. One example we have discovered is mu-crystallin which is not only a major lens protein in some mammals but is also expressed abundantly in retina where it may serve as an enzyme of amino acid metabolism with importance for photoreceptors. We have cloned and mapped the human gene for mu-crystallin and are examining its expression in retina and lens. Another major group of lens proteins, the gamma-crystallins were thought to be highly specific for lens, but we have found that gammas-crystallin is also expressed in retina. We have cloned and mapped the mouse gammas gene and shown it to be the locus for Opj, a genetic cataract in mice. We have synthesized normal and Opj mutant gammas proteins to study the basis of the cataract. We have also synthesized proteins for structural analysis of AIM1, a melanoma-associated protein related to gamma-crystallin. We have characterized the gene promoter of zeta-crystallin and have shown that a specific form of Pax6 is essential for its function. A second factor, Nrl, is required for high level expression and a third element, designated BPE, is necessary to suppress "collateral" expression in brain. Macrophage migration inhibitory factor (MIF) is a delayed early response gene expressed in lens, retina, cornea and elsewhere. We have cloned and mapped human and mouse genes for MIF and the related D-dopachrome tautomerase. We have synthesized recombinant proteins to study MIF activity in suppression of NK T-cell cytotoxicity in aqueous humor, the possible enzymatic role of MIF and the structure of MIF in solution. A collaborative "gene knockout" experiment for MIF is also in progress.

正常的透镜依赖于丰富的蛋白质，称为晶体蛋白， 它的清晰度。晶体蛋白是多功能蛋白质， 但通常保留非透镜中重要的其他功能 组织中我们发现的一个例子是mu-crystallin， 在某些哺乳动物中仅是一种主要的透镜蛋白， 在视网膜中大量存在，可作为氨基酸酶 新陈代谢对感光细胞很重要。 我们已经克隆和 绘制了人类μ晶体蛋白基因的图谱， 在视网膜和透镜中表达。另一组主要的透镜蛋白， γ-晶体蛋白被认为对透镜具有高度特异性， 但我们发现γ-晶状体蛋白也在视网膜中表达。 我们已经克隆并绘制了小鼠gammas基因， Opj基因座，一种小鼠遗传性白内障。我们已经合成 正常和Opj突变γ蛋白研究的基础上， 白内障我们还合成了蛋白质用于结构分析 AIM 1是一种与γ-晶体蛋白相关的黑色素瘤相关蛋白。 我们已经表征了zeta-晶体蛋白的基因启动子， 表明Pax 6的特定形式对其功能至关重要。一 第二因子Nrl是高水平表达所需的，第三因子Nrl是高水平表达所需的。 指定为BPE的元素对于抑制“抵押品”是必要的 在大脑中的表达。 巨噬细胞移动抑制因子（MIF）是一种迟发性的早期免疫抑制因子。 在透镜、视网膜、角膜和其他地方表达的应答基因。我们 已经克隆并绘制了人类和小鼠的MIF基因及其相关的 多巴色素互变异构酶我们已经合成了重组蛋白， 巨噬细胞移动抑制因子抑制NK细胞杀伤活性研究 水状体，MIF的可能的酶促作用和结构 在溶液中。一项合作的“基因敲除”实验， 多国临时部队也在进行中。