PMID: 8938714Jan 1, 1996Paper

Localization and interaction of epitope-tagged GIRK1 and CIR inward rectifier K+ channel subunits

Neuropharmacology
M E KennedyD E Clapham

Abstract

GIRK1 and CIR are G-protein activated inward rectifier K+ channel subunits that combine to form the heteromultimer IKACh, the G beta gamma-activated atrial K channel responsible for the vagal slowing of heart rate. Epitope-tagged channel subunits were constructed by the introduction of distinct six amino acid epitopes into the C-termini or putative extracellular domains of GIRK1 and CIR. Carboxyl-terminal tagged subunits were activated by purified G beta gamma subunits in inside-out patches when expressed in Cos cells. Interestingly, insertion of three amino acids into the putative extracellular domain of GIRK1 resulted in an inactive subunit that acted as a dominant negative subunit when coexpressed with wild type GIRK1 and CIR in Xenopus oocytes. The epitope-tagged CIR-AU1 subunit coimmunoprecipitated GIRK1-AU5 from metabolically labeled Cos cells. Immunofluorescence labeling of Cos cells localized GIRK1-AU5 to internal cytoskeletal structures that co-stained with antibodies against the intermediate filament protein, vimentin. CIR-AU1 localized primarily to the plasma membrane. Double immunofluorescence labeling showed that GIRK1-AU5 plasma membrane staining was detectable only when coexpressed with CIR-AU1.

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Citations

Jun 23, 2001·Journal of the American College of Cardiology·P KovoorD E Clapham
Aug 2, 2008·Journal of Cardiovascular Pharmacology·Joachim R Ehrlich
Jul 22, 1999·Annals of the New York Academy of Sciences·K WickmanD E Clapham
Sep 18, 2010·The European Journal of Neuroscience·Francisco CiruelaRafael Luján
Feb 15, 2005·American Journal of Transplantation : Official Journal of the American Society of Transplantation and the American Society of Transplant Surgeons·Lev KoyrakhKevin Wickman
Dec 20, 2007·Journal of Neurochemistry·Carolina AguadoRafael Luján
Apr 3, 2009·Biochemical and Biophysical Research Communications·Keiko IshiharaYoshihiro Kubo
Jan 21, 2018·Scientific Reports·Allison AndersonKevin Wickman
Jan 21, 2010·Physiological Reviews·Hiroshi HibinoYoshihisa Kurachi
Jan 16, 1999·The Journal of Biological Chemistry·M E KennedyD E Clapham
Jan 20, 2001·The Journal of Biological Chemistry·S Corey, D E Clapham
Apr 18, 1997·The Journal of Biological Chemistry·R WoodwardR D Murrell-Lagnado
Jun 27, 1998·The Journal of Biological Chemistry·G KrapivinskyD E Clapham
Oct 9, 1998·The Journal of Biological Chemistry·S Corey, D E Clapham
Jul 19, 2008·Molecular Biology Reports·Xiongtao LiuQiangsun Zheng
Mar 28, 1998·The Journal of Biological Chemistry·S CoreyD E Clapham
May 11, 2017·Scientific Reports·Ezequiel Marron Fernandez de VelascoKevin Wickman
May 15, 2001·American Journal of Physiology. Lung Cellular and Molecular Physiology·E D MichelakisS L Archer
Jan 3, 2004·The Journal of Biological Chemistry·Tooraj Mirshahi, Diomedes E Logothetis
Jan 1, 1997·Annual Review of Pharmacology and Toxicology·D E Clapham, E J Neer

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