Membrane Cholesterol Strongly Influences Confined Diffusion of Prestin

dc.citation.firstpage1627
dc.citation.journalTitleBiophysical Journal
dc.citation.lastpage1636
dc.citation.volumeNumber103
dc.contributor.authorKamar, R.I.
dc.contributor.authorOrgan-Darling, L.E.
dc.contributor.authorRaphael, R.M.
dc.date.accessioned2013-01-09T19:44:03Z
dc.date.available2013-01-09T19:44:03Z
dc.date.issued2012-10
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.
dc.embargo.termsnone
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.
dc.identifier.doihttp://dx.doi.org/10.1016/j.bpj.2012.07.052
dc.identifier.urihttps://hdl.handle.net/1911/68535
dc.language.isoeng
dc.publisherBiophysical Society
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.
dc.titleMembrane Cholesterol Strongly Influences Confined Diffusion of Prestin
dc.typeJournal article
dc.type.dcmiText
dc.type.publicationpublisher version
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