Identification and characterization of proteins bound to the cytoplasmic domain of protocadherins

与几种原钙粘蛋白的胞质结构域相关的蛋白质的鉴定和表征已经进行。候选人的结合蛋白进行了分离，通过亲和层析使用GST融合蛋白的细胞质结构域的原钙粘素，并通过质谱法进行了鉴定。利用酵母双杂交技术分离结合蛋白。结果表明，候选蛋白主要属于细胞骨架蛋白。然后，进行免疫荧光显微镜和免疫沉淀，以显示候选蛋白实际上与原钙粘蛋白在体内。初步结果显示，在几个案件中有希望。然而，一个主要的障碍是，在大多数情况下，没有针对候选人的良好抗体。我们现在正在生产针对候选人的抗体，以方便实验。我们还计划对候选人进行异位表达实验，以阐明原钙粘蛋白的生物学作用。

Identification and characterization of proteins that associate with the cytoplasmic domains of several protocadherins have been carried out. Candidate binding proteins were isolated by affinity chromatography using GST-fusion proteins of the cytoplasmic domains of protocadherins and were identified by mass-spectroscopy. Yeast two-hybrid method was also used to isolate the binding proteins. The results revealed candidate proteins mostly belonged to cytoskeletal proteins. Immuno-fluorescence microscopy and immunoprecipitation were, then, performed to show that the candidate proteins actually associated with protocadherins in vivo. Preliminary results showed promissing in several cases. However, a major obstacle is that good antibodies against the candidates are not available in most cases. We are now producing the antibodies against the candidates' to facilitate the experiments. We are also planning ectopic expression experiments of the candidates to clarify the biological role of protocadherins.