Label-free cell phenotypic profiling decodes the composition and signaling of an endogenous ATP-sensitive potassium channel

Haiyan Sun, Ying Wei, Huayun Deng, Qiaojie Xiong, Min Li, Joydeep Lahiri, Ye Fang

Research output: Contribution to journalArticlepeer-review

Abstract

Current 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 KATP blocker profiling showed that the pinacidil DMR is due to the activation of SUR2/Kir6.2 KATP channels in HepG2C3A cells. Kinase inhibition and RNAi knockdown showed that the pinacidil activated KATP channels 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.

Original languageEnglish (US)
Article number4934
JournalScientific reports
Volume4
DOIs
StatePublished - May 12 2014

ASJC Scopus subject areas

  • General

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