Identification of inner ear specific protein binds to connexin 26

鉴定与连接蛋白 26 结合的内耳特异性蛋白

• 批准号: 18591876
• 负责人: KOKAI Yasuo
• 金额: $ 2.49万
• 依托单位: Sapporo Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2006
• 资助国家: 日本
• 起止时间: 2006 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18591876/
• 关键词: inner ear; proteome; impaired hearing; neuroscinece; biological molecule; 神経科学; プロテーム

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.

据报道，连接蛋白26在听力障碍中起着重要作用，尽管其分子机制仍然鲜为人知。为了研究连接蛋白26在听力障碍中的作用，我们开发了表达人缔合蛋白26的转基因小鼠。在这些转基因小鼠中，表达了所有检查的人connexin26。使用抗FLAG抗体拉下与连接蛋白26结合的蛋白。通过飞行类型质谱时间分析结合的蛋白质，以使这些结合的蛋白质中蛋白质谱。比较这些器官之间的结合蛋白，仅在内耳的提取物中检测到几种蛋白质。除了耳朵特异性蛋白外，我们还检测到每个器官特异性蛋白。我们目前正在纯化这些蛋白质以鉴定氨基酸序列。总体而言，这种方法非常有效地在理论上与任何类型的蛋白质鉴定有器官特定的结合蛋白。

项目成果

血漿のプロテオミクス

血浆蛋白质组学

• 发表时间: 2007
• 期刊: 日本臨床免疫学会誌 30(3)
• 作者: 苗代 康可;ら
• 通讯作者: ら

移植片対宿主疾患の検査法

移植物抗宿主病检测方法

• 发表时间: 2007