Identification of inner ear specific protein binds to connexin 26

据报道，缝隙连接蛋白26在听力障碍中起着重要作用，但其分子机制尚不清楚。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，人类连接蛋白26在所有被检测的器官中都有表达，这表明连接蛋白26普遍表达。从内耳、肝脏、小肠、肾脏和脾中提取蛋白质。抗FLAG抗体被用来下调连接蛋白26结合的蛋白。用飞行时间质谱仪对结合蛋白进行分析，得到结合蛋白的蛋白质图谱。比较这些器官的结合蛋白，只在内耳提取液中检测到几种蛋白。除了耳朵特有的蛋白质外，我们还检测到了每个器官的特有蛋白质。我们目前正在提纯这些蛋白质，以鉴定氨基酸序列。总的来说，这种方法被发现非常有效地识别器官特异性结合蛋白，理论上是任何类型的蛋白。

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.