TRANSMEMBRANE ASSEMBLAGE OF THE PHOTORECEPTOR CONNECTING CILIUM AND MOTILE CILIUM TRANSITION ZONE CONTAIN A COMMON IMMUNOLOGICAL EPITOPE
TRANSMEMBRANE ASSEMBLAGE OF THE PHOTORECEPTOR CONNECTING CILIUM AND MOTILE CILIUM TRANSITION ZONE CONTAIN A COMMON IMMUNOLOGICAL EPITOPE
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DOI：
10.1002/cm.970170408
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发表时间：
1990-01-01
影响因子：
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通讯作者：
BESHARSE, JC
中科院分区：
文献类型：
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作者：
HORST, CJ;JOHNSON, LV;BESHARSE, JC
连接纤毛的光感受器具有独特的跨膜组装，其稳定地将细胞表面糖缀合物与下面的轴丝细胞骨架连接。光感受器连接纤毛和运动纤毛的过渡区之间的结构相似性表明，这种组合也可能存在于运动纤毛。使用富含洗涤剂提取的光感受器轴丝的亚细胞部分，三个高分子量的糖缀合物（425，600和700 kD）先前被确定为潜在的组件的组合。通过寡糖特性和结合的特异性单克隆抗体，我们已经验证了本地化的425 kD的糖缀合物的跨膜组装。结合凝集素花生凝集素（PNA）的硝酸纤维素印迹上的425 kD的糖缀合物，和分离的洗涤剂提取的轴丝，进行了评估后处理的酶神经氨酸酶和O-聚糖酶。与425 kD糖缀合物结合的变化精确地抑制了与完整轴丝结合的变化，支持了425 kD糖缀合物是跨膜组装的组分的假设。此外，结果表明，425 kD的糖缀合物含有唾液酸化的半乳糖-N-乙酰半乳糖胺寡糖，其O-连接到蛋白质骨架上。为了直接评估425 kD糖缀合物的分布，我们制备了针对该糖缀合物的单克隆抗体。抗体K26仅识别轴丝部分的transblots上的425 kD。K26免疫反应完整的轴索是相同的PNA染色所看到的。分离的光感受器和整个视网膜的K26染色独特地定位于连接纤毛的区域。因此，在光感受器中，425 kD不仅是跨膜组装的组分，而且完全限于连接纤毛。基于形态学上的相似性，光感受器连接纤毛被认为是同源的运动纤毛的过渡区。因此，我们用K26单克隆抗体对输卵管上皮进行染色。免疫反应性仅限于运动纤毛基部的过渡区区域。这表明，光感受器连接纤毛和运动纤毛过渡区是免疫相关的。
The photoreceptor connecting cilium bears a unique transmembrane assemblage which stably links cell surface glycoconjugates with the underlying axonemal cytoskeleton. Structural similarities between the photoreceptor connecting cilium and the transition zone of motile cilia suggests that this assemblage may also be present in motile cilia. Using a subcellular fraction enriched in detergent-extracted photoreceptor axonemes, three high molecular mass glycoconjugates (425, 600, and 700 kD) were previously identified as potential components of the assemblage. Through oligosaccharide characterization and binding of a specific monoclonal antibody, we have verified the localization of the 425 kD glycoconjugate to the transmembrane assemblage. Binding of the lectin peanut agglutinin (PNA) to the 425 kD glycoconjugate on nitrocellulose blots, and to isolated detergent-extracted axonemes, was assessed following treatment with the enzymes neuraminidase and O-glycanase. Changes in binding to the 425 kD glycoconjugate precisely paralleled changes in binding to intact axonemes, supporting the hypothesis that the 425 kD glycoconjugate is a component of the transmembrane assemblage. Furthermore, the results suggest that the 425 kD glycoconjugate contains sialated galactose-N-acetylgalactosamine oligosaccharides which are O-linked to the protein backbone. To directly assess the distribution of the 425 kD glycoconjugate, we produced a monoclonal antibody directed against this glycoconjugate. The antibody, K26, recognizes only the 425 kD on transblots of the axoneme fraction. K26 immunoreactivity of intact axonemes is identical to that seen by PNA staining. K26 staining of isolated photoreceptors and whole retina is uniquely localized to the region of the connecting cilium. Thus, in the photoreceptor, the 425 kD is not only a component of the transmembrane assemblage but is also completely restricted to the connecting cilium.Based on morphological similarities, the photoreceptor connecting cilium is thought to be homologous to the transition zone of the motile cilium. As such, we have stained oviduct epithelium with the K26 monoclonal antibody. Immunoreactivity is restricted to the region of the transition zone at the base of motile cilia. This demonstrates that the photoreceptor connecting cilium and motile cilium transition zone are immunologically related.