Prestin topology: localization of protein epitopes in relation to the plasma membrane
Prestin topology: localization of protein epitopes in relation to the plasma membrane
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DOI：
10.1097/00001756-200107030-00032
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发表时间：
2001-07
期刊：
影响因子：
1.7
通讯作者：
Jing Zheng;Kevin B. Long;W. Shen;L. Madison;P. Dallos
中科院分区：
文献类型：
--
作者：
Jing Zheng;Kevin B. Long;W. Shen;L. Madison;P. Dallos
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 transfec- tion 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.
