TY - JOUR TI - PDZD7-MYO7A complex identified in enriched stereocilia membranes AU - Morgan, Clive P AU - Krey, Jocelyn F AU - Grati, M'hamed AU - Zhao, Bo AU - Fallen, Shannon AU - Kannan-Sundhari, Abhiraami AU - Liu, Xue Zhong AU - Choi, Dongseok AU - Müller, Ulrich AU - Barr-Gillespie, Peter G A2 - Nathans, Jeremy VL - 5 PY - 2016 DA - 2016/08/15 SP - e18312 C1 - eLife 2016;5:e18312 DO - 10.7554/eLife.18312 UR - https://doi.org/10.7554/eLife.18312 AB - While more than 70 genes have been linked to deafness, most of which are expressed in mechanosensory hair cells of the inner ear, a challenge has been to link these genes into molecular pathways. One example is Myo7a (myosin VIIA), in which deafness mutations affect the development and function of the mechanically sensitive stereocilia of hair cells. We describe here a procedure for the isolation of low-abundance protein complexes from stereocilia membrane fractions. Using this procedure, combined with identification and quantitation of proteins with mass spectrometry, we demonstrate that MYO7A forms a complex with PDZD7, a paralog of USH1C and DFNB31. MYO7A and PDZD7 interact in tissue-culture cells, and co-localize to the ankle-link region of stereocilia in wild-type but not Myo7a mutant mice. Our data thus describe a new paradigm for the interrogation of low-abundance protein complexes in hair cell stereocilia and establish an unanticipated link between MYO7A and PDZD7. KW - hair bundle KW - auditory KW - vestibular KW - mass spectrometry KW - immunoaffinity JF - eLife SN - 2050-084X PB - eLife Sciences Publications, Ltd ER -