STRUCTURAL FEATURES OF KERATIN AND RELATED INTERMEDIATE FILAMENTS

角蛋白中间丝的结构、功能及表达 人类皮肤和其他细胞类型的相关中间丝， 正在接受调查。我们已经构建了一系列合成的 与角蛋白链序列相对应的多肽参与 组装细丝中的重要重叠区域，对于生物物理 结构研究，并用于X射线结晶学。其中几个 这些合成肽已经被注射到活细胞中，以探索 它们的动态行为。其中一些用作非常特异的试剂。 对于所有类型的中间长丝的断裂，或仅对于 角蛋白细丝。为了探索END结构域的功能 序列，我们使用了双酵母杂交系统。在其他研究中，我们 已经确定了类型II的V1亚区的一部分 角蛋白参与了与其他蛋白质的交联性角化 细胞包膜，这一过程似乎协调了结构 角蛋白细丝细胞骨架与细胞周缘的组织 在终末分化角质形成细胞中。其他交联剂的分析 揭示角蛋白是附着在桥粒蛋白成分上的 桥粒通过一系列相关的中间丝间接形成 相关蛋白质。我们已经生产了一种永生细胞系，使用 1例表皮松解症患者的表皮角质形成细胞 角化过度伴角蛋白10基因突变的基因研究 治疗策略。

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.