ArticleThe Journal of cell biology1991
High-purity isolation of bullfrog hair bundles and subcellular and topological localization of constituent proteins.
Article in The Journal of cell biology, 1991. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 37 papers.
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Who cites it
37 citing papers in PubMed, 103 citations in OpenAlex.
- Electron cryo-tomography of vestibular hair-cell stereocilia.Journal of structural biology · 2019Article
- Analysis of the Proteome of Hair-Cell Stereocilia by Mass Spectrometry.Methods in enzymology · 2017Article
- Plastin 1 widens stereocilia by transforming actin filament packing from hexagonal to liquid.The Journal of cell biology · 2016Article
- Neuroplastin Isoform Np55 Is Expressed in the Stereocilia of Outer Hair Cells and Required for Normal Outer Hair Cell Function.The Journal of neuroscience : the official journal of the Society for Neuroscience · 2016Article
- Article
- Annexin A5 is the Most Abundant Membrane-Associated Protein in Stereocilia but is Dispensable for Hair-Bundle Development and Function.Scientific reports · 2016Article
- Stereocilia-staircase spacing is influenced by myosin III motors and their cargos espin-1 and espin-like.Nature communications · 2016Article
- Hair-bundle proteomes of avian and mammalian inner-ear utricles.Scientific data · 2015Article
- Assembly of hair bundles, an amazing problem for cell biology.Molecular biology of the cell · 2015Review
- Gene Expression by Mouse Inner Ear Hair Cells during Development.The Journal of neuroscience : the official journal of the Society for Neuroscience · 2015Article
- Article
- The proteome of mouse vestibular hair bundles over development.Scientific data · 2015Article
- Correlation of actin crosslinker and capper expression levels with stereocilia growth phases.Molecular & cellular proteomics : MCP · 2014Article
- Molecular architecture of the chick vestibular hair bundle.Nature neuroscience · 2013Article
- Large membrane domains in hair bundles specify spatially constricted radixin activation.The Journal of neuroscience : the official journal of the Society for Neuroscience · 2012Article
- Divalent counterions tether membrane-bound carbohydrates to promote the cohesion of auditory hair bundles.Biophysical journal · 2011Article
- The R109H variant of fascin-2, a developmentally regulated actin crosslinker in hair-cell stereocilia, underlies early-onset hearing loss of DBA/2J mice.The Journal of neuroscience : the official journal of the Society for Neuroscience · 2010Article
- Twinfilin 2 regulates actin filament lengths in cochlear stereocilia.The Journal of neuroscience : the official journal of the Society for Neuroscience · 2009Article
- Article
- The chloride intracellular channel protein CLIC5 is expressed at high levels in hair cell stereocilia and is essential for normal inner ear function.The Journal of neuroscience : the official journal of the Society for Neuroscience · 2006Article
Corrections and comments
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Authors and funding
2 authors at 1 institution in 1 country.
Funding
Abstract
The small number of hair cells in auditory and vestibular organs severely impedes the biochemical characterization of the proteins involved in mechano-electrical transduction. By developing an efficient and clean "twist-off" method of hair bundle isolation, and by devising a sensitive, nonradioactive method to detect minute quantities of protein, we have partially overcome this limitation and have extensively classified the proteins of the bundles. To isolate hair bundles, we glue the saccular macula of the bullfrog to a glass coverslip, expose the tissue to a molten agarose solution, and allow the agarose to solidify to a firm gel. By rotating the gel disk with respect to the fixed macula, we isolate the hair bundles by shearing them at their mechanically weak bases. The plasma membranes of at least 80% of the stereocilia reseal. To visualize the proteins of the hair bundle, we covalently label them with biotin, separate them by SDS-PAGE, and transfer them to a charged nylon membrane. We can detect less than 500 fg of protein by probing the membrane with streptavidin-alkaline phosphatase and detecting the chemiluminescent product from the hydrolysis of the substrate 3-(4-methoxyspiro-(1,2-dioxetane-3,2'-tricyclo-[3.3.1. 1(3.7)]decan)-4-yl) phenyl phosphate (AMPPD). These techniques reveal a distinct constellation of proteins in and associated with hair bundles. Several proteins, such as calmodulin, calbindin, actin, tubulin, and fimbrin, have previously been described. A second class of proteins in the preparation appears to be derived from extracellular sources. Finally, several heretofore undescribed bundle proteins are identified and characterized by their membrane topology, subcellular localization, and glycosidase and protease sensitivities.
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