Open AccessFunctional expression and microdomain localization of prestin in cultured cells
Author(s) -
Sturm Angela K.,
Rajagopalan Lavanya,
Yoo Donald,
Brownell William E.,
Pereira Fred A.
Publication year - 2007
Publication title -
otolaryngology–head and neck surgery
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.232
H-Index - 121
eISSN - 1097-6817
pISSN - 0194-5998
DOI - 10.1016/j.otohns.2006.10.030
Subject(s) - prestin , microbiology and biotechnology , colocalization , chemistry , lipid microdomain , lipid raft , intracellular , transfection , hek 293 cells , motor protein , membrane protein , membrane , biophysics , biology , biochemistry , gene , microtubule
Prestin is an essential component of the molecular motor of cochlear outer hair cells that contribute to frequency selectivity and sensitivity of mammalian hearing. A model system to study prestin employs its transfection into cultured HEK 293 cells. Our goal was to characterize prestin's trafficking pathway and localization in the plasma membrane. Methods We used immuno‐colocalization of prestin with intracellular and plasma membrane markers and sucrose density fractionation to analyze prestin in membrane compartments. Voltage clamping was used to measure nonlinear capacitance (NLC), prestin's electrical signature. Results & Discussion Prestin targets to the membrane by 24 hours post‐transfection when NLC is measurable. Prestin then concentrates into membrane foci that colocalize and fractionate with membrane microdomains. Depleting membrane cholesterol content altered prestin localization and NLC. Conclusion Prestin activity in HEK 293 cells results from expression in the plasma membrane and altering membrane lipid content affects prestin localization and activity.