EXPRESSION, STRUCTURE AND FUNCTION OF FILAGGRIN

丝聚蛋白的表达、结构和功能

• 批准号: 3810937
• 负责人: P STEINERT
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF ARTHRITIS AND MUSCULOSKELETAL AND SKIN DISEASES
• 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/3810937
• 关键词: cell differentiation; gene expression; genetic disorder; genetically modified animals; human tissue; intermediate filaments; keratin; laboratory mouse; molecular cloning; nuclear magnetic resonance spectroscopy; pathologic keratinization; protein sequence; proteolysis; skin

Filaggrin is a major differentiation product of terminally differentiating mammalian epidermal cells, that is thought to be involved in the aggregation and specific alignment of keratin intermediate filaments during the final stages of differentiation. Thus filaggrin is an important example of an intermediate filament-associated protein. We have isolated both cDNA and genomic clones which show that filaggrin is initially expressed as a large polyprotein precursor, filaggrin, that is subsequently proteolytically processed into individual functional filaggrin molecules. In the human system, the precursor consists of 3 allelic size variants, containing 10, 11 or 12 tandem repeats that segregated by normal Mendelian genetic mechanisms. In addition, these repeats show considerable sequence variation, so that any two repeats are only about 85% homologous to each other; so far, we find that of the 324 amino acid residues of the filaggrin repeats, about 40% of the positions are variable. We have constructed genomic fragments for the production of transgenic mice. We have begun a systematic analysis of regulatory sequences that control the expression of this gene system. We are studying the method of interaction of filaggrin with keratin intermediate filaments by use of solid state NMR techniques. Since there are a number of keratinizing disorders of the skin for which there is some evidence involving incorrect expression of the profilaggrin gene, we have begun a systematic search for the possible role of filaggrin in genetic diseases of keratinization.

聚丝蛋白是终末分化的主要分化产物， 哺乳动物的表皮细胞，被认为参与了 角蛋白中间丝的聚集和特异性排列 分化的最后阶段。因此丝聚蛋白是一个重要的例子 一种中间毒性相关蛋白质。我们分离出了两个cDNA 以及显示聚丝蛋白最初表达为聚丝蛋白的基因组克隆， 大的多聚蛋白前体，聚丝蛋白，随后 蛋白水解加工成单独的功能性聚丝蛋白分子。 在人类系统中，前体由3个等位基因大小变体组成， 含有10、11或12个串联重复序列，这些重复序列被正常孟德尔遗传分离， 遗传机制。此外，这些重复序列显示出相当大的序列 变异，因此任何两个重复序列仅与每个重复序列约85%同源。 其他;到目前为止，我们发现丝聚蛋白的324个氨基酸残基中 重复，约40%的位置是可变的。我们已经构建 用于生产转基因小鼠的基因组片段。我们已经开始了 系统分析控制表达的调控序列， 这个基因系统。我们正在研究丝聚蛋白相互作用的方法 与角蛋白中间丝通过使用固态NMR技术。 由于存在许多皮肤的角质化病症， 有一些证据涉及前丝聚蛋白的不正确表达 基因，我们已经开始了对丝聚蛋白可能的作用进行系统的研究 角化的遗传性疾病。