Project Details
Identification of hair bundle proteins from sensory hair cells using mass spectrometry
Applicant
Dr. Jung-Bum Shin, Ph.D.
Subject Area
Molecular Biology and Physiology of Neurons and Glial Cells
Term
from 2004 to 2006
Project identifier
Deutsche Forschungsgemeinschaft (DFG) - Project number 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 Programme
Research Fellowships
International Connection
USA
Cooperation Partner
Professor Dr. Peter G. Gillespie