HEK cell MYO7A-PDZD7 immunoprecipitation

Through its activity in stereocilia, MYO7A (myosin VIIA) is essential for hair cell function in the inner ear. Utilizing multiple stages of immunoaffinity enrichment, we have developed a strategy that allows us to partially purify stereocilia membranes from thousands of chick inner ears and isolate low-abundance MYO7A protein complexes from those membranes, including one with the scaffolding protein PDZD7. To confirm the interaction of PDZD7 and MYO7A, we expressed them singly or together in HEK cells, and immunoprecipitated detergent-soluble fractions with specific antibodies. PDZD7 and MYO7A interacted strongly in these tissue-culture cells.