Enhancement of TREK1 channel surface expression by protein-protein interaction with beta-COP.
TREK1 belongs to a family of two-pore-domain K(+) (K(2P)) channels and produce background currents that regulate cell excitability. In the present study, we identified a vesicle transport protein, beta-COP, as an interacting partner by yeast two-hybrid screening of a human brain cDNA library with N-terminal region of TREK1 (TREK1-N) as bait. Several in vitro and in vivo binding assays confirmed the protein-protein interaction between TREK1 and beta-COP. We also found that beta-COP was associated with TREK1 in native condition at the PC3 cells. When RFP-beta-COP was co-transfected with GFP-TREK1 into COS-7 cells, both proteins were found localized to the plasma membrane. In addition, the channel activity and surface expression of GFP-TREK1 increased dramatically by co-transfection with RFP-beta-COP. Surface expression of the TREK1 channel was also clearly reduced with the addition of beta-COP-specific shRNA. Collectively, these data suggest that beta-COP plays a critical role in the forward transport of TREK1 channel to the plasma membrane.
Kim
Eunju
E
Department of Physiology, Gyeongsang National University School of Medicine, Jinju 660-751, South Korea.
Hwang
Eun Mi
EM
Yarishkin
Oleg
O
Yoo
Jae Cheal
JC
Kim
Donggyu
D
Park
Nammi
N
Cho
Minhee
M
Lee
Young Sun
YS
Sun
Choong-Hyun
CH
Yi
Gwan-Su
GS
Yoo
Jiyun
J
Kang
Dawon
D
Han
Jaehee
J
Hong
Seong-Geun
SG
Park
Jae-Yong
JY
D000818:Animals; D019556:COS Cells; D002462:Cell Membrane / Q000378:metabolism*; D002522:Chlorocebus aethiops; D020756:Coatomer Protein / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D024683:Potassium Channels, Tandem Pore Domain / Q000235:genetics / Q000378:metabolism*; D025941:Protein Interaction Mapping; D020798:Two-Hybrid System Techniques
D020756:Coatomer Protein; D024683:Potassium Channels, Tandem Pore Domain; C103742:potassium channel protein TREK-1