The dynamics of epiplakin null keratinocytes and the production of transgenic mice of epiplakin

Epiplakin是一种细胞骨架连接蛋白，最初是从一名表皮下水泡病患者的血清中鉴定为自身抗原。为了检测表膜蛋白与中间丝的结合能力，我们用包含表膜蛋白不同结构域和亚结构域的融合蛋白进行了狭缝印迹分析。在表位蛋白B区的4.6个拷贝中，至少有两个是融合蛋白与角蛋白结合所必需的。在这些检测中，连接结构域的重复结构也在表膜蛋白与角蛋白的结合中发挥了重要作用，同时也增加了表膜蛋白连接结构域中的重复结构，这参与了与IF结合的增加。还检测到与波形蛋白和结蛋白的结合类似但较弱的结合。这些观察表明，外膜蛋白在B结构域和连接器域的高度重复结构是该分子在蛋白家族中的独特特征，在该分子的功能中发挥着重要作用。

AbstractEpiplakin, a cytoskeletal linker protein, was originally identified as an autoantigen in a serum spec men obtained from a patient with subepidermal blistering disease. To examine the binding ability of epiplakin with intermediate filaments (IFs), we performed slot-blot assays using fusion proteins that included various domains and subdomains of epiplakin. At least two of the 4.6 copies in the B domains of epiplakin were necessary for the binding of fusion proteins to keratin. The repeated structures of linker domains also played an important role in the binding of epiplakin to keratin in these assays while also increasing the repeated structure in the linker domain of epiplakin which is involved in the increased binding to IFs. A similar but weaker binding to vimentin and desmin was also detected. These observations indicated that the highly repeated structures of epiplakin in both the B and the linker domains, which is the unique feature of this molecule in the plakin family, play an essential role in the functioning of this molecule.