Label-free cell phenotypic profiling decodes the composition and signaling of an endogenous ATP-sensitive potassium channel
0301 basic medicine
572
Drug Evaluation, Preclinical
Gene Expression
Ligands
Article
Cell Line
Membrane Potentials
03 medical and health sciences
KATP Channels
cell signaling
Cluster Analysis
Humans
RNA, Messenger
Protein Kinase Inhibitors
rho-Associated Kinases
0303 health sciences
Pinacidil
Janus Kinase 3
Janus Kinase 2
Actins
Phenotype
Gene Knockdown Techniques
ion channel
RNA Interference
Protein Kinases
signal transduction
Signal Transduction
DOI:
10.1038/srep04934
Publication Date:
2014-05-12T09:06:57Z
AUTHORS (7)
ABSTRACT
AbstractCurrent technologies for studying ion channels are fundamentally limited because of their inability to functionally link ion channel activity to cellular pathways. Herein, we report the use of label-free cell phenotypic profiling to decode the composition and signaling of an endogenous ATP-sensitive potassium ion channel (KATP) in HepG2C3A, a hepatocellular carcinoma cell line. Label-free cell phenotypic agonist profiling showed that pinacidil triggered characteristically similar dynamic mass redistribution (DMR) signals in A431, A549, HT29 and HepG2C3A, but not in HepG2 cells. Reverse transcriptase PCR, RNAi knockdown and KATPblocker profiling showed that the pinacidil DMR is due to the activation of SUR2/Kir6.2 KATPchannels in HepG2C3A cells. Kinase inhibition and RNAi knockdown showed that the pinacidil activated KATPchannels trigger signaling through Rho kinase and Janus kinase-3 and cause actin remodeling. The results are the first demonstration of a label-free methodology to characterize the composition and signaling of an endogenous ATP-sensitive potassium ion channel.
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