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Identification of the large-conductance background K+ channel in mouse B cells as TREK-2.

Authors
 Haifeng Zheng  ;  Joo Hyun Nam  ;  Bo Pang  ;  Dong Hoon Shin  ;  Ji Seon Kim  ;  Yang-Sook Chun  ;  Jong-Wan Park  ;  Hyowon Bang  ;  Woo Kyung Kim  ;  Yung E. Earm  ;  Sung Joon Kim 
Citation
 AMERICAN JOURNAL OF PHYSIOLOGY-CELL PHYSIOLOGY, Vol.297(1) : 188-197, 2009 
Journal Title
 AMERICAN JOURNAL OF PHYSIOLOGY-CELL PHYSIOLOGY 
ISSN
 0363-6143 
Issue Date
2009
MeSH
1-Phosphatidylinositol 4-Kinase/metabolism ; Adenosine Triphosphate/metabolism ; Animals ; Arachidonic Acid/metabolism ; B-Lymphocytes/drug effects ; B-Lymphocytes/enzymology ; B-Lymphocytes/metabolism* ; Calcium/metabolism ; Cell Line ; Cell Shape ; Cloning, Molecular ; Humans ; Hydrogen-Ion Concentration ; Mechanotransduction, Cellular ; Membrane Potentials ; Mice ; Phosphatidylinositol 4,5-Diphosphate/metabolism ; Phosphodiesterase Inhibitors/pharmacology ; Potassium/metabolism* ; Potassium Channels, Tandem Pore Domain/drug effects ; Potassium Channels, Tandem Pore Domain/genetics ; Potassium Channels, Tandem Pore Domain/metabolism* ; Protein Kinase Inhibitors/pharmacology ; RNA Interference ; Rats ; Transfection ; Type C Phospholipases/metabolism
Keywords
K2P channel ; arachidonic acid ; PI kinase ; membrane stretch ; immune cells
Abstract
Mouse B cells and their cell line (WEHI-231) express large-conductance background K(+) channels (LK(bg)) that are activated by arachidonic acids, characteristics similar to TREK-2. However, there is no evidence to identify the molecular nature of LK(bg); some properties of LK(bg) were partly different from the reported results of TREK type channels. In this study, we compared the properties of cloned TREK-2 and LK(bg) in terms of their sensitivities to ATP, phosphatidylinositol 4,5-bisphosphate (PIP(2)), intracellular pH (pH(i)), and membrane stretch. Similar to the previous findings of LK(bg), TREK-2 showed spontaneous activation after membrane excision (i-o patch) and were inhibited by MgATP or by PIP(2). The inhibition by MgATP was prevented by wortmannin, suggesting membrane-delimited regulation of TREKs by phosphoinositide (PI) kinase. The same was observed with the property of LK(bg); the activation of TREK-2 by membrane stretch was suppressed by U73122 (PLC inhibitor). As with the known properties of TREK-2, LK(bg) were activated by acidic pH(i) and inhibited by PKC activator. Finally, we confirmed the expression of TREK-2 in WEHI-231 by using RT-PCR and immunoblot analyses. The amplitude of background K(+) current and the TREK-2 expression in WEHI-231 were commonly decreased by genetic knockdown of TREK-2 using small interfering RNA. The downregulation of TREK-2 attenuated Ca(2+)-influx induced by arachidonic acid in WEHI-231. As a whole, these results strongly indicate that TREK-2 encodes LK(bg) in mouse B cells. We also newly suggest that the low activity of TREK-2 in intact cells is due to the inhibition by intrinsic PIP(2).
Files in This Item:
T200905263.pdf Download
DOI
10.1152/ajpcell.00052.2009
Appears in Collections:
5. Research Institutes (연구소) > Research Center for Human Natural Defense System (생체방어연구센터) > 1. Journal Papers
Yonsei Authors
Nam, Joo Hyun(남주현)
URI
https://ir.ymlib.yonsei.ac.kr/handle/22282913/105801
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