title: Label-free cell phenotypic profiling decodes the composition and signaling of an endogenous ATP-sensitive potassium channel.
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.
authors.affiliation: 1] Biochemical Technologies, Science and Technology Division, Corning Incorporated, Corning, NY 14831, United States of America [2].
authors.forename: Haiyan
authors.initials: H
authors.lastname: Sun
authors.affiliation: Biochemical Technologies, Science and Technology Division, Corning Incorporated, Corning, NY 14831, United States of America.
authors.forename: Ying
authors.initials: Y
authors.lastname: Wei
authors.affiliation: Biochemical Technologies, Science and Technology Division, Corning Incorporated, Corning, NY 14831, United States of America.
authors.forename: Huayun
authors.initials: H
authors.lastname: Deng
authors.affiliation: 1] The Solomon H. Snyder Department of Neuroscience and High Throughput Biology Center, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, United States of America [2].
authors.forename: Qiaojie
authors.initials: Q
authors.lastname: Xiong
authors.affiliation: The Solomon H. Snyder Department of Neuroscience and High Throughput Biology Center, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, United States of America.
authors.forename: Min
authors.initials: M
authors.lastname: Li
authors.affiliation: Biochemical Technologies, Science and Technology Division, Corning Incorporated, Corning, NY 14831, United States of America.
authors.forename: Joydeep
authors.initials: J
authors.lastname: Lahiri
authors.affiliation: Biochemical Technologies, Science and Technology Division, Corning Incorporated, Corning, NY 14831, United States of America.
authors.forename: Ye
authors.initials: Y
authors.lastname: Fang
mesh_terms: D000199:Actins / Q000378:metabolism; D002460:Cell Line; D016000:Cluster Analysis; D004353:Drug Evaluation, Preclinical / Q000379:methods; D015870:Gene Expression; D055785:Gene Knockdown Techniques; D006801:Humans; D053614:Janus Kinase 2 / Q000378:metabolism; D053616:Janus Kinase 3 / Q000378:metabolism; D054086:KATP Channels / Q000819:agonists / Q000235:genetics / Q000378:metabolism*; D008024:Ligands; D008564:Membrane Potentials / Q000187:drug effects; D010641:Phenotype; D020110:Pinacidil / Q000494:pharmacology; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D011494:Protein Kinases / Q000378:metabolism; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics; D015398:Signal Transduction; D054460:rho-Associated Kinases / Q000378:metabolism
chemical_list: D000199:Actins; D054086:KATP Channels; D008024:Ligands; D047428:Protein Kinase Inhibitors; D012333:RNA, Messenger; D020110:Pinacidil; D011494:Protein Kinases; D053614:Janus Kinase 2; D053616:Janus Kinase 3; D054460:rho-Associated Kinases