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Article: Characterization of recombinant human cardiac KCNQ1/KCNE1 channels (I Ks) stably expressed in HEK 293 cells

TitleCharacterization of recombinant human cardiac KCNQ1/KCNE1 channels (I Ks) stably expressed in HEK 293 cells
Authors
Issue Date2006
PublisherSpringer New York LLC. The Journal's web site is located at http://link.springer.de/link/service/journals/00232/
Citation
Journal Of Membrane Biology, 2006, v. 210 n. 3, p. 183-192 How to Cite?
AbstractThe present study was designed to characterize pharmacological, biophysical and electrophysiological properties of the recombinant human cardiac I Ks (KCNQ1/KCNE1) channels at physiological temperature. Human cardiac KCNQ1 and KCNE1 genes were cotransfected into HEK 293 cells, and a cell clone stably expressing both genes was selected. Membrane currents were recorded using a perforated patch-clamp technique. The typical I Ks was slowly activated upon depolarization voltages in HEK 293 cells stably expressing human cardiac KCNQ1 and KCNE1 genes, and the current was inhibited by I Ks blockers HMR 1556 and chromanol 293B, with 50% inhibitory concentrations (IC 50s) of 83.8 nM and 9.2 μM, respectively. I Ks showed a significant temperature-dependent increase in its magnitude upon elevating bath temperature to 36°C from room temperature (21°C). The current was upregulated by the β-adrenoceptor agonist isoproterenol, and the effect was reversed by H89. In addition, I Ks was inhibited by Ba 2+ in a concentration-dependent manner (IC 50 = 1.4 mM). Action potential clamp revealed a "bell-shaped" time course of I Ks during the action potential, and maximal peak current was seen at the plateau of the action potential. A significant use- and frequency-dependent increase of I Ks was observed during a train of action potential clamp. These results indicate that the recombinant human cardiac I Ks stably expressed in HEK 293 cells is similar to native I Ks in drug sensitivity and regulated by Ba 2+ and β-adrenoceptor via the cyclic adenosine monophosphate/protein kinase A pathway. Importantly, the current exhibits significant temperature dependence, a bell-shaped time course during action potential and prominent use- or frequency-dependent accumulation during a train of action potentials. © Springer Science+Business Media, Inc. 2006.
Persistent Identifierhttp://hdl.handle.net/10722/163013
ISSN
2015 Impact Factor: 1.991
2015 SCImago Journal Rankings: 0.738
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorDong, MQen_US
dc.contributor.authorLau, CPen_US
dc.contributor.authorGao, Zen_US
dc.contributor.authorTseng, GNen_US
dc.contributor.authorLi, GRen_US
dc.date.accessioned2012-09-05T05:26:31Z-
dc.date.available2012-09-05T05:26:31Z-
dc.date.issued2006en_US
dc.identifier.citationJournal Of Membrane Biology, 2006, v. 210 n. 3, p. 183-192en_US
dc.identifier.issn0022-2631en_US
dc.identifier.urihttp://hdl.handle.net/10722/163013-
dc.description.abstractThe present study was designed to characterize pharmacological, biophysical and electrophysiological properties of the recombinant human cardiac I Ks (KCNQ1/KCNE1) channels at physiological temperature. Human cardiac KCNQ1 and KCNE1 genes were cotransfected into HEK 293 cells, and a cell clone stably expressing both genes was selected. Membrane currents were recorded using a perforated patch-clamp technique. The typical I Ks was slowly activated upon depolarization voltages in HEK 293 cells stably expressing human cardiac KCNQ1 and KCNE1 genes, and the current was inhibited by I Ks blockers HMR 1556 and chromanol 293B, with 50% inhibitory concentrations (IC 50s) of 83.8 nM and 9.2 μM, respectively. I Ks showed a significant temperature-dependent increase in its magnitude upon elevating bath temperature to 36°C from room temperature (21°C). The current was upregulated by the β-adrenoceptor agonist isoproterenol, and the effect was reversed by H89. In addition, I Ks was inhibited by Ba 2+ in a concentration-dependent manner (IC 50 = 1.4 mM). Action potential clamp revealed a "bell-shaped" time course of I Ks during the action potential, and maximal peak current was seen at the plateau of the action potential. A significant use- and frequency-dependent increase of I Ks was observed during a train of action potential clamp. These results indicate that the recombinant human cardiac I Ks stably expressed in HEK 293 cells is similar to native I Ks in drug sensitivity and regulated by Ba 2+ and β-adrenoceptor via the cyclic adenosine monophosphate/protein kinase A pathway. Importantly, the current exhibits significant temperature dependence, a bell-shaped time course during action potential and prominent use- or frequency-dependent accumulation during a train of action potentials. © Springer Science+Business Media, Inc. 2006.en_US
dc.languageengen_US
dc.publisherSpringer New York LLC. The Journal's web site is located at http://link.springer.de/link/service/journals/00232/en_US
dc.relation.ispartofJournal of Membrane Biologyen_US
dc.subject.meshAction Potentials - Drug Effects - Physiologyen_US
dc.subject.meshAdrenergic Beta-Agonists - Pharmacologyen_US
dc.subject.meshBarium - Pharmacologyen_US
dc.subject.meshCells, Cultureden_US
dc.subject.meshChromans - Pharmacologyen_US
dc.subject.meshCyclic Amp-Dependent Protein Kinases - Antagonists & Inhibitorsen_US
dc.subject.meshGene Expression Regulation - Physiologyen_US
dc.subject.meshHumansen_US
dc.subject.meshIsoproterenol - Pharmacologyen_US
dc.subject.meshIsoquinolines - Pharmacologyen_US
dc.subject.meshKcnq1 Potassium Channel - Antagonists & Inhibitors - Physiologyen_US
dc.subject.meshKidney - Embryology - Metabolismen_US
dc.subject.meshMembrane Potentials - Drug Effectsen_US
dc.subject.meshPatch-Clamp Techniquesen_US
dc.subject.meshPotassium Channel Blockers - Pharmacologyen_US
dc.subject.meshPotassium Channels, Voltage-Gated - Antagonists & Inhibitors - Physiologyen_US
dc.subject.meshProtein Kinase Inhibitors - Pharmacologyen_US
dc.subject.meshRecombinant Proteins - Metabolismen_US
dc.subject.meshSulfonamides - Pharmacologyen_US
dc.subject.meshTransfectionen_US
dc.titleCharacterization of recombinant human cardiac KCNQ1/KCNE1 channels (I Ks) stably expressed in HEK 293 cellsen_US
dc.typeArticleen_US
dc.identifier.emailLi, GR:grli@hkucc.hku.hken_US
dc.identifier.authorityLi, GR=rp00476en_US
dc.description.naturelink_to_subscribed_fulltexten_US
dc.identifier.doi10.1007/s00232-006-0006-5en_US
dc.identifier.pmid16909339-
dc.identifier.scopuseid_2-s2.0-33748104214en_US
dc.identifier.hkuros122230-
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-33748104214&selection=ref&src=s&origin=recordpageen_US
dc.identifier.volume210en_US
dc.identifier.issue3en_US
dc.identifier.spage183en_US
dc.identifier.epage192en_US
dc.identifier.isiWOS:000240198400003-
dc.publisher.placeUnited Statesen_US
dc.identifier.scopusauthoridDong, MQ=7202127303en_US
dc.identifier.scopusauthoridLau, CP=7401968501en_US
dc.identifier.scopusauthoridGao, Z=16549711200en_US
dc.identifier.scopusauthoridTseng, GN=7006468947en_US
dc.identifier.scopusauthoridLi, GR=7408462932en_US

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