Figure 5. Identification and quantitation of FSCN2 in chicken vestibular stereocilia. a , Structure of FSCN2 protein. Colors indicate identity of residues in three-way comparison of chicken, mouse, and Xenopus FSCN2. Peptides used for antibody generation are indicated by black bars and antibody numbers; the position of the mutation in DBA/2J is indicated with an asterisk. The positions of the four β-trefoil domains are indicated below. b , Representative MS2 spectrum of a FSCN2 peptide identified by tandem mass spectrometry from a chicken hair bundle preparation. The intensity of peptide fragments, typically fragmented at peptide bonds during collision-induced dissociation, is plotted against the mass-to-charge ratio (m/z). Note the nearly complete y- and b-ion ladders, representing fragmentation sequentially at successive peptide bonds from the N and C termini. c , Distribution of FSCN2 peptides identified by X! Tandem algorithm from hair bundles. The width of each bar indicates the length of the identified peptide, the position of the bar along the x-axis corresponds to its position in the FSCN2 sequence, the height of the bar indicates the number of identical peptides identified, and the color of the bar corresponds to log(e), the statistical significance for peptide identification, averaged over all identical peptides. Note that the peptide analyzed in b was one of those represented by the black bar at ∼380 residues. d , Crosslinker/actin abundance ratios in E20 chick and P5 rat vestibular hair bundles. e , Immunoblot detection of FSCN2 in purified hair bundles with antibody 36. Bu, 100 ng of total hair-bundle protein; Ag, amount of agarose equivalent to that in bundles lane; Ut, 2 μg of peeled utricular epithelium; BP, 2 μg of peeled basilar papilla epithelium; Re, 1.5 μg of retina; and Br, 2 μg of total brain protein. f , g , Profile scans of bundle and utricular epithelium lanes from e .
Image published in: Shin JB et al. (2010)
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