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: 6160820
• 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/6160820
• 关键词: X ray crystallography; cell differentiation; conformation; crosslink; cytoskeletal proteins; human tissue; intercellular connection; intermediate filaments; keratin; keratinocyte; protein sequence; protein structure function; skin; synthetic peptide; tissue /cell culture; yeast two hybrid system

The structure, function and expression of keratin intermediate filaments of human skin, and the related intermediate filaments of other cell types, are being investigated. While the roles of these proteins in many genetic diseases are now well understood, further structural studies are necessary to develop rational approaches to therapy. We have constructed a series of synthetic peptides of sequence corresponding to 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. We have expressed keratins 5 and 14 to explore their structures and functions in vivo. 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 of stratified squamous epithelia, a process which coordinates 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.

角蛋白中间丝的结构、功能和表达 以及其他细胞的相关中间纤维 类型，正在调查中。 虽然这些蛋白质在许多疾病中的作用 遗传疾病现在已经很好地了解，进一步的结构研究是 需要制定合理的治疗方法。 我们已经构建 一系列合成肽，其序列对应于角蛋白 在组装的重要重叠区域中涉及的链 生物物理结构研究用和X射线用灯丝 结晶学 其中几种合成肽已经被注射到 活细胞中探索它们的动态行为。 其中一些 作为非常特异性的试剂，用于破坏所有类型的 中间丝，或仅角蛋白丝。 为了 为了探索末端结构域序列的功能， 混合系统我们表达了角蛋白5和14，以探索它们的生物学活性。 结构和功能。 在其他研究中，我们已经确定 II型角蛋白的V1亚结构域区域的一部分参与 在与其他蛋白质交联的皮质细胞包膜， 复层鳞状上皮，一个协调结构的过程， 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