Identification of inner ear specific protein binds to connexin 26

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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.