Study of phototransduction micromachinary on the disk membrane

外节盘的形成过程贯穿于生物体的整个生命周期，为外节组件的持续更新提供了机制。在具有动态结构的圆盘膜上，存在大量与光转导相关的蛋白质，如视紫红质，当片段接收到光时，转导级联发生作用。由于级联反应进展非常快，因此可以预测积聚蛋白质的微机械的形成。免疫细胞化学显示微管蛋白α、β、γ和tau在盘状膜上共定位。为了鉴定外段的微管蛋白特异性结合蛋白，我们使用了免疫沉淀和质谱法。用质谱法鉴定了部分蛋白。tau蛋白和膜联蛋白II与微管蛋白和微管蛋白共同沉淀。这些结果表明微管蛋白在光导微机械中起支架的作用。由于牛的BSE无法获得牛视网膜，我们将材料改为青蛙视网膜。制备了青蛙活性氧馏分，并采用2D-PAGE和质谱法对其进行了分析。对136个凝胶点进行分析，鉴定出35种蛋白质。14个蛋白为视紫红质，其中14个蛋白起源于外节段，含有视紫红质、磷酸二酯酶α和β、转导蛋白α和β、阻滞蛋白和s-调节蛋白。18个蛋白起源于内节段，含有HSC70、烯醇化酶、GTP-rho结合蛋白、神经元中间丝和ATP合成酶。肌动蛋白和微管蛋白是两者共有的。

The process of outer segment disk formation occurs throughout the life of an organism, providing a mechanism for continual renewal of outer segment components. On a disk membrane with dynamic structure, a lot of proteins related to phototransduction, such as rhodopsin, exist and the transduction cascade works when the segment has received light. Since the cascade reaction progresses very quickly, formation of micromachinery that accumulates the proteins can be predicted. Immunocytochemistry revealed that tubulin alpha, beta, gamma and tau were co-localized on the disk membrane. To identify the tubulin specific binding proteins in outer segments, we demonstrated immunoprecipitation and mass spectrometry. Some proteins were identified by mass spectrometry. The tau and an annexin II were co-precipitated with tubulin beta and gamma. These results suggest that the tubulin plays the role of the scaffold of a phototransduction micromachinary. Since the bovine retina stopped obtaining for the BSE of the bovine, we changed material into the frog retina. We prepared frog ROS fraction and analyzed the fraction by 2D-PAGE and mass spectrometry. Thirty five proteins had been identified when 136-gel spots were analyzed. Forteen proteins were rhodopsin, The proteins of 14 originated in outer segment and contained rhodopsin, phosphodiesterase alpha and beta, transducin alpha and beta, arrestin and s-modulin. The proteins of 18 originated in inner segment and contained HSC70,enolase, GTP-rho binding protein, neuronal intermediate filament and ATP synthase. The actin, and a tubulin alpha and beta were common to both.