Analysis of the Proteome of Hair-Cell Stereocilia by Mass Spectrometry

Research output: Chapter in Book/Report/Conference proceedingChapter

2 Citations (Scopus)

Abstract

Characterization of proteins that mediate mechanotransduction by hair cells, the sensory cells of the inner ear, is hampered by the scarcity of these cells and their sensory organelle, the hair bundle. Mass spectrometry, with its high sensitivity and identification precision, is the ideal method for determining which proteins are present in bundles and what proteins they interact with. We describe here the isolation of mouse hair bundles, as well as preparation of bundle protein samples for mass spectrometry. We also describe protocols for data-dependent (shotgun) and parallel reaction monitoring (targeted) mass spectrometry that allow us to identify and quantify proteins of the hair bundle. These sensitive methods are particularly useful for comparing proteomes of wild-type mice and mice with deafness mutations affecting hair-bundle proteins.

Original languageEnglish (US)
Title of host publicationMethods in Enzymology
PublisherAcademic Press Inc.
Pages329-354
Number of pages26
Volume585
DOIs
StatePublished - 2017

Publication series

NameMethods in Enzymology
Volume585
ISSN (Print)00766879
ISSN (Electronic)15577988

Fingerprint

Stereocilia
Proteome
Hair
Mass spectrometry
Mass Spectrometry
Proteins
Firearms
Deafness
Inner Ear
Organelles
Cells
Mutation
Monitoring

Keywords

  • Actin
  • Cytoskeleton
  • Espin
  • Hair bundles
  • Hair cells
  • Ion trap
  • Orbitrap
  • Shotgun
  • Stereocilia
  • Targeted

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Cite this

Krey, J. F., Wilmarth, P., David, L., & Barr-Gillespie, P. (2017). Analysis of the Proteome of Hair-Cell Stereocilia by Mass Spectrometry. In Methods in Enzymology (Vol. 585, pp. 329-354). (Methods in Enzymology; Vol. 585). Academic Press Inc.. https://doi.org/10.1016/bs.mie.2016.09.023

Analysis of the Proteome of Hair-Cell Stereocilia by Mass Spectrometry. / Krey, J. F.; Wilmarth, Phillip; David, Larry; Barr-Gillespie, Peter.

Methods in Enzymology. Vol. 585 Academic Press Inc., 2017. p. 329-354 (Methods in Enzymology; Vol. 585).

Research output: Chapter in Book/Report/Conference proceedingChapter

Krey, JF, Wilmarth, P, David, L & Barr-Gillespie, P 2017, Analysis of the Proteome of Hair-Cell Stereocilia by Mass Spectrometry. in Methods in Enzymology. vol. 585, Methods in Enzymology, vol. 585, Academic Press Inc., pp. 329-354. https://doi.org/10.1016/bs.mie.2016.09.023
Krey JF, Wilmarth P, David L, Barr-Gillespie P. Analysis of the Proteome of Hair-Cell Stereocilia by Mass Spectrometry. In Methods in Enzymology. Vol. 585. Academic Press Inc. 2017. p. 329-354. (Methods in Enzymology). https://doi.org/10.1016/bs.mie.2016.09.023
Krey, J. F. ; Wilmarth, Phillip ; David, Larry ; Barr-Gillespie, Peter. / Analysis of the Proteome of Hair-Cell Stereocilia by Mass Spectrometry. Methods in Enzymology. Vol. 585 Academic Press Inc., 2017. pp. 329-354 (Methods in Enzymology).
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