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

点击翻译按钮获取中文摘要

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.