期刊
NEUROREPORT
卷 12, 期 9, 页码 1929-1935出版社
LIPPINCOTT WILLIAMS & WILKINS
DOI: 10.1097/00001756-200107030-00032
关键词
anion transporters; DRA; outer hair cells; PDS; prestin; SLC26A6; topology
Computer modeling of the outer hair cell (OHC) motor protein prestin produces ambiguous results regarding transmembrane regions and localization of its termini. To determine the location of prestin's N- and C-termini, we created prestin constructs with synthetic epitopes located immediately upstream or downstream of prestin. The spatial distribution of these epitopes was studied in prestin-transfected cells using immunofluorescence. In permeabilized cells, antibodies label the plasma membrane of 30% of the cells, reflecting transfection efficiency. Under non-permeabilizing conditions, the few labeled cells also displayed a lack of plasma membrane integrity. These data suggest that prestin's N-and C-termini are cytoplasmic. Furthermore, prestin staining in OHCs was observed only under permeabilizing conditions. These results implicate prestin's N- and C-termini as portions that may interact with other cytoplasmic proteins. A model of prestin membrane topology is also considered based on the results. NeuroReport 12:1929-1935 (C) 2001 Lippincott Williams & Wilkins.
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