Identification of hair bundle proteins from sensory hair cells using mass spectrometry

• Antragsteller: Dr. Jung-Bum Shin, Ph.D.
• Fachliche Zuordnung: Molekulare Biologie und Physiologie von Nerven- und Gliazellen
• Förderung: Förderung von 2004 bis 2006
• Projektkennung: Deutsche Forschungsgemeinschaft (DFG) - Projektnummer 5429842

Hair cells are the sensory organs of the auditory and vestibular system, enabling us to sense sound and balance. The transduction apparatus, which transduces mechanical stimuli into electrical signals, resides in the hair bundle, protrusions of the hair cell apical membrane. Although a method for isolating hair bundles exists, the limited sensitivity of previous protein sequencing methods precluded a comprehensive identification of the extremely scarce components of the hair bundle. However, recent developments in proteomic technologies have become sensitive enough to allow direct sequencing of hair bundle proteins. In this study, I plan to use the isotope-coded affinity tag (ICAT) method, in combination with mass spectrometry (MS), to identify hair bundle components on the protein level. I will label mouse hair bundles with one ICAT reagent and bundle-removed sensory epithelia with a similar reagent of slightly larger mass, fractionate labeled peptides and identify peptides derived from proteins enriches in hair bundles. Those peptides will be sequenced using mass spectrometry and the original proteins will be identidfied. The localisation of interesting proteins in hair bundles will be confirmed by protein interaction studies and electrophysiological recording from hair cell cultures will teach us more about the function of these proteins.

• DFG-Verfahren: Forschungsstipendien
• Internationaler Bezug: USA
• Kooperationspartner: Professor Dr. Peter G. Gillespie