Effect of Rab5 on large-conductance Ca2+-actived K+ channels (BKca) expressed in HEK293 Cells
ZHOU Dan
LIU Xueru
LI Tao
LIU Li
TAN Xiaoqiu
LIU Yulin
Abstract：Objective To investigate the effect of Rab5 on large-conductance Ca2+-activated K+ channels (BKCa channels) expressed in HEK293 cells.Methods HEK293 cells in the logarithmic phase were randomly divided to four groups:the control group,Rab5WT group,Rab5CA group and Rab5DN group.HEK293 cells were transfected with FlaghSlo-GFP plasmid in each group and with pcDNA3.1 for control group,Rab5WT for Rab5WT group,Rab5CA for Rab5CA group and Rab5DN for Rab5DN group,respectively.Total 4 μg plasmid with 1∶1 to each plasmid was transfected into 3.5 mm dish.Transfection rate was observed under fluorescence microscopy and the effect of Rab5 on BKCa channels was detected by_patch clamp technique,Western blotting and flow cytometry when the trasfection rate was above 70％.Results Compared with the control group,Rab5 WT and Rab5 CA significantly increased BKCa macroscopic currents,while BKCa currents were decreased in the Rab5DN group.Rab5 WT and Rab5 CA increased the expression of membranous BKCa protein in HEK293 cells,while Rab5DN decreased the expression (all P ＜0.05).Rab5 WT and Rab5 CA increased the Flag+/GFP+ ratio (all P＜0.05),while Rab5DN decreased the ratio (P ＜0.05).Conclusion Rab5 significantly increases BKCa currents and channel expression on cell surface and may promote the transport process of BKCa currents to cell membrane in HEK293 cells.
Keywords：small G protein familylarge-conductance Ca2+-activated K+ channelsprotein transportgene transfection
Publication Date：2017-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：5( 21-24,后插1 )