Identification of inner ear specific protein binds to connexin 26

连接蛋白26在听力损伤中起重要作用，但其分子机制仍不清楚。为了研究连接蛋白26在听力损伤中的作用，我们开发了表达人连接蛋白26的转基因小鼠。在这些转基因小鼠中，人连接蛋白26在所有检查的器官中表达，表明连接蛋白26的普遍表达。从内耳、肝脏、小肠、肾脏和脾脏制备蛋白质提取物。抗FLAG抗体用于拉下与连接蛋白26结合的蛋白质。通过飞行时间型质谱法分析结合的蛋白质，以在这些结合的蛋白质中形成蛋白质谱。比较各器官的结合蛋白，有几种蛋白仅在内耳提取液中检测到。除了耳特异性蛋白外，我们还检测了各器官特异性蛋白。我们目前正在纯化这些蛋白质以鉴定氨基酸序列。总的来说，发现这种方法对于鉴定理论上任何类型蛋白的器官特异性结合蛋白非常有效。

Connexin 26 has been reported to play an important role for hearing impairment, though its molecular mechanism is still poorly understood. To study a role of connexin 26 in hearing impairment, we developed transgenic mice expressing human connexin 26. In these transgenic mice, human connexin26 was expressed all the organs examined, suggesting universal expression of connexin 26. Protein extracts were prepared from inner ear, liver, small intestine, kidney and spleen. Anti-FLAG antibody was used to pull down proteins bound to connexin 26. Bound proteins were analyzed by time of flight type mass spectrometry to make protein profile in these bound proteins. Comparing bound proteins among these organs, several proteins were only detedted in extracts from inner ear. In addition to ear specific proteins, we also detected each organ specific proteins. We are currently purifying these proteins to identify amino acid sequences. Collectively, this approach is found to be very effective to identify organ specific binding proteins to theoretically any type of proteins.