Abstract |
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.
|
Authors | Clive P Morgan, Jocelyn F Krey, M'hamed Grati, Bo Zhao, Shannon Fallen, Abhiraami Kannan-Sundhari, Xue Zhong Liu, Dongseok Choi, Ulrich Müller, Peter G Barr-Gillespie |
Journal | eLife
(Elife)
Vol. 5
(08 15 2016)
ISSN: 2050-084X [Electronic] England |
PMID | 27525485
(Publication Type: Journal Article)
|
Chemical References |
- Carrier Proteins
- Myo7a protein, mouse
- Myosin VIIa
- PDZD7 protein, mouse
- Myosins
|
Topics |
- Animals
- Carrier Proteins
(analysis, isolation & purification)
- Mass Spectrometry
- Membranes
(chemistry)
- Mice
- Myosin VIIa
- Myosins
(analysis, isolation & purification)
- Protein Binding
- Stereocilia
(chemistry)
|