Membrane Cholesterol Strongly Influences Confined Diffusion of Prestin

dc.citation.firstpage1627en_US
dc.citation.journalTitleBiophysical Journalen_US
dc.citation.lastpage1636en_US
dc.citation.volumeNumber103en_US
dc.contributor.authorKamar, R.I.en_US
dc.contributor.authorOrgan-Darling, L.E.en_US
dc.contributor.authorRaphael, R.M.en_US
dc.date.accessioned2013-01-09T19:44:03Zen_US
dc.date.available2013-01-09T19:44:03Zen_US
dc.date.issued2012-10en_US
dc.description.abstractPrestin is the membrane motor protein that drives outer hair cell (OHC) electromotility, a process that is essential for mammalian hearing. Prestin function is sensitive to membrane cholesterol levels, and numerous studies have suggested that prestin localizes in cholesterol-rich membrane microdomains. Previously, fluorescence recovery after photobleaching experiments were performed in HEK cells expressing prestin-GFP after cholesterol manipulations, and revealed evidence of transient confinement. To further characterize this apparent confined diffusion of prestin, we conjugated prestin to a photostable fluorophore (tetramethylrhodamine) and performed single-molecule fluorescence microscopy. Using single-particle tracking, we determined the microscopic diffusion coefficient from the full time course of the mean-squared deviation. Our results indicate that prestin undergoes diffusion in confinement regions, and that depletion of membrane cholesterol increases confinement size and decreases confinement strength. By interpreting the data in terms of a mathematical model of hop-diffusion, we quantified these cholesterol-induced changes in membrane organization. A complementary analysis of the distribution of squared displacements confirmed that cholesterol depletion reduces prestin confinement. These findings support the hypothesis that prestin function is intimately linked to membrane organization, and further promote a regulatory role for cholesterol in OHC and auditory function.en_US
dc.embargo.termsnoneen_US
dc.identifier.citationKamar, R.I., Organ-Darling, L.E. and Raphael, R.M.. "Membrane Cholesterol Strongly Influences Confined Diffusion of Prestin." <i>Biophysical Journal,</i> 103, (2012) Biophysical Society: 1627-1636. http://dx.doi.org/10.1016/j.bpj.2012.07.052.en_US
dc.identifier.doihttp://dx.doi.org/10.1016/j.bpj.2012.07.052en_US
dc.identifier.urihttps://hdl.handle.net/1911/68535en_US
dc.language.isoengen_US
dc.publisherBiophysical Societyen_US
dc.rightsArticle is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use.en_US
dc.titleMembrane Cholesterol Strongly Influences Confined Diffusion of Prestinen_US
dc.typeJournal articleen_US
dc.type.dcmiTexten_US
dc.type.publicationpublisher versionen_US
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