PMID- 25148687
OWN - NLM
STAT- MEDLINE
DCOM- 20150206
LR  - 20211021
IS  - 1083-351X (Electronic)
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 289
IP  - 41
DP  - 2014 Oct 10
TI  - Tandem pore domain halothane-inhibited K+ channel subunits THIK1 and THIK2 
      assemble and form active channels.
PG  - 28202-12
LID - 10.1074/jbc.M114.600437 [doi]
AB  - Despite a high level of sequence homology, tandem pore domain halothane-inhibited 
      K(+) channel 1 (THIK1) produces background K(+) currents, whereas THIK2 is 
      silent. This lack of activity is due to a unique combination of intracellular 
      retention and weak basal activity in the plasma membrane. Here, we designed THIK 
      subunits containing dominant negative mutations (THIK1(DN) and THIK2(DN)). 
      THIK2(DN) mutant inhibits THIK1 currents, whereas THIK1(DN) inhibits an activated 
      form of THIK2 (THIK2-A155P-I158D). In situ proximity ligation assays and 
      Forster/fluorescence resonance energy transfer (FRET) experiments support a 
      physical association between THIK1 and THIK2. Next, we expressed covalent tandems 
      of THIK proteins to obtain expression of pure heterodimers. Td-THIK1-THIK2 (where 
      Td indicates tandem) produces K(+) currents of amplitude similar to 
      Td-THIK1-THIK1 but with a noticeable difference in the current kinetics. Unlike 
      Td-THIK2-THIK2 that is mainly detected in the endoplasmic reticulum, 
      Td-THIK1-THIK2 distributes at the plasma membrane, indicating that THIK1 can mask 
      the endoplasmic reticulum retention/retrieval motif of THIK2. Kinetics and 
      unitary conductance of Td-THIK1-THIK2 are intermediate between THIK1 and THIK2. 
      Altogether, these results show that THIK1 and THIK2 form active heteromeric 
      channels, further expanding the known repertoire of K(+) channels.
CI  - (c) 2014 by The American Society for Biochemistry and Molecular Biology, Inc.
FAU - Blin, Sandy
AU  - Blin S
AD  - From LabEx ICST, Institut de Pharmacologie Moleculaire et Cellulaire, CNRS, and 
      Universite de Nice Sophia Antipolis, 660 Route des Lucioles, 06560 Valbonne, 
      France and.
FAU - Chatelain, Franck C
AU  - Chatelain FC
AD  - From LabEx ICST, Institut de Pharmacologie Moleculaire et Cellulaire, CNRS, and 
      Universite de Nice Sophia Antipolis, 660 Route des Lucioles, 06560 Valbonne, 
      France and.
FAU - Feliciangeli, Sylvain
AU  - Feliciangeli S
AD  - From LabEx ICST, Institut de Pharmacologie Moleculaire et Cellulaire, CNRS, and 
      Universite de Nice Sophia Antipolis, 660 Route des Lucioles, 06560 Valbonne, 
      France and.
FAU - Kang, Dawon
AU  - Kang D
AD  - the Department of Physiology and Institute of Health Sciences, Gyeongsang 
      National University, School of Medicine, Jinju 660-751, South Korea.
FAU - Lesage, Florian
AU  - Lesage F
AD  - From LabEx ICST, Institut de Pharmacologie Moleculaire et Cellulaire, CNRS, and 
      Universite de Nice Sophia Antipolis, 660 Route des Lucioles, 06560 Valbonne, 
      France and lesage@ipmc.cnrs.fr.
FAU - Bichet, Delphine
AU  - Bichet D
AD  - From LabEx ICST, Institut de Pharmacologie Moleculaire et Cellulaire, CNRS, and 
      Universite de Nice Sophia Antipolis, 660 Route des Lucioles, 06560 Valbonne, 
      France and.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20140822
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Chlorides)
RN  - 0 (KCNK12 protein, human)
RN  - 0 (KCNK13 protein, human)
RN  - 0 (Potassium Channels, Tandem Pore Domain)
RN  - 0 (Recombinant Proteins)
RN  - RWP5GA015D (Potassium)
SB  - IM
MH  - Amino Acid Sequence
MH  - Animals
MH  - Chlorides/*chemistry/metabolism
MH  - Dogs
MH  - Gene Expression Regulation
MH  - HEK293 Cells
MH  - Humans
MH  - Ion Transport
MH  - Kinetics
MH  - Madin Darby Canine Kidney Cells
MH  - Membrane Potentials
MH  - Models, Molecular
MH  - Molecular Sequence Data
MH  - Mutation
MH  - Oocytes/cytology/physiology
MH  - Patch-Clamp Techniques
MH  - Potassium/*chemistry/metabolism
MH  - Potassium Channels, Tandem Pore Domain/*chemistry/genetics/metabolism
MH  - Protein Structure, Secondary
MH  - Protein Structure, Tertiary
MH  - Recombinant Proteins/chemistry/genetics/metabolism
MH  - Xenopus laevis
PMC - PMC4192476
OTO - NOTNLM
OT  - Electrophysiology
OT  - K2P Channels
OT  - KCNK
OT  - Plasma Membrane
OT  - Potassium Channel
OT  - Protein Assembly
OT  - Protein Complex
EDAT- 2014/08/26 06:00
MHDA- 2015/02/07 06:00
PMCR- 2015/10/10
CRDT- 2014/08/23 06:00
PHST- 2014/08/23 06:00 [entrez]
PHST- 2014/08/26 06:00 [pubmed]
PHST- 2015/02/07 06:00 [medline]
PHST- 2015/10/10 00:00 [pmc-release]
AID - S0021-9258(20)37081-2 [pii]
AID - M114.600437 [pii]
AID - 10.1074/jbc.M114.600437 [doi]
PST - ppublish
SO  - J Biol Chem. 2014 Oct 10;289(41):28202-12. doi: 10.1074/jbc.M114.600437. Epub 
      2014 Aug 22.