Dunia I, Recouvreur M, Nicolas P, Kumar N, Bloemendal H, Benedetti E L
Institut J. Monod CNRS, Université Paris VII, France.
J Cell Sci. 1998 Aug;111 ( Pt 15):2109-20. doi: 10.1242/jcs.111.15.2109.
The SDS-fracture immunolabeling technique, unlike conventional freeze-fracture, provides direct evidence for the biochemical nature of membrane constituents. SDS-fracture immunolabeling shows that during differentiation of lens fiber cells the onset of junctional assembly is characterized by the presence of small clusters and linear arrays comprising connexins alpha3 and alpha8. At this initial stage MP26, a major fiber membrane constituent, appears to be colocalized with these two connexins. The application of double-immunogold labeling reveals that when large junctional plaques are assembled MP26 becomes mainly associated with the periphery of the junctional domains. This type of distribution suggests that MP26 may play a role in the clustering and gathering of connexons. In aged nuclear fiber membranes connexins, MP26 and their proteolytic derivatives form an orthogonal lattice of repeating subunits.
与传统的冷冻断裂不同，SDS断裂免疫标记技术为膜成分的生化性质提供了直接证据。SDS断裂免疫标记显示，在晶状体纤维细胞分化过程中，连接组装的开始表现为存在由连接蛋白α3和α8组成的小簇和线性阵列。在这个初始阶段，主要的纤维膜成分MP26似乎与这两种连接蛋白共定位。双重免疫金标记的应用表明，当大型连接斑组装时，MP26主要与连接域的周边相关联。这种分布类型表明MP26可能在连接子的聚集和集合中起作用。在老年核纤维膜中，连接蛋白、MP26及其蛋白水解衍生物形成重复亚基的正交晶格。
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