STRUCTURAL FEATURES OF KERATIN AND RELATED INTERMEDIATE FILAMENTS

角蛋白及相关中间丝的结构特征

• 批准号: 2568362
• 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/2568362
• 关键词: X ray crystallography; cell differentiation; cell line; conformation; congenital ichthyosis; crosslink; cytoskeletal proteins; gene mutation; gene therapy; human tissue; intercellular connection; intermediate filaments; keratin; keratinocyte; protein sequence; protein structure function; skin; synthetic peptide; tissue /cell culture; yeasts

The structure, function and expression of keratin intermediate filaments of human skin, and the related intermediate filaments of other cell types, are being investigated. We have constructed a series of synthetic peptides of sequence corresponding keratin chains which are involved in the important overlap regions in assembled filaments, for biophysical structural studies, and for use for X-ray crystallography. Several of these synthetic peptides have been injected into living cells to explore their dynamic behavior. Some of these function as very specific reagents for the disruption of all types of intermediate filaments, or of only keratin filaments. In order to explore the functions of end domain sequences, we have used the two yeast hybrid system. In other studies, we have determined that a portion of the V1 subdomain region of type II keratins is involved in crosslinking with other proteins to the cornified cell envelope, a process which seems to coordinate the structural organization of the keratin filament cytoskeleton with the cell periphery in terminally differentiated keratinocytes. Analyses of other crosslinks reveal that the keratins are attached to the desmoplakin component of desmosomes indirectly through a series of related intermediate filament associated proteins. We have produced an immortalized cell line using epidermal keratinocytes obtained from a patient with epidermolytic hyperkeratosis with a keratin 10 gene mutation in order to explore gene therapy strategies.

角蛋白中间丝的结构、功能和表达 以及其他细胞类型的相关中间纤维， 正在接受调查 我们构建了一系列的合成 肽序列对应的角蛋白链，涉及 组装细丝中的重要重叠区域， 结构研究，并用于X射线晶体学。 几 这些合成肽被注入活细胞中， 动态行为。 其中一些作为非常特殊的试剂 用于破坏所有类型的中间丝，或仅用于破坏 角蛋白丝 为了探讨端结构域的功能 序列，我们使用了两个酵母杂交系统。 在其他研究中，我们 已经确定II型V1子域区域的一部分 角蛋白参与与其它蛋白质的交联 细胞包膜，一个似乎协调结构的过程， 角蛋白丝细胞骨架与细胞周边的组织 在终末分化的角质形成细胞中。 其他交联分析 显示角蛋白附着于桥粒斑蛋白成分， 桥粒通过一系列相关的中间丝间接连接 相关蛋白质 我们已经生产了一种永生化细胞系， 表皮角质形成细胞从患有表皮炎的患者获得， 角化过度伴角蛋白10基因突变，以探索基因 治疗策略。