Functional expression of an epitope-tagged G protein-coupled K+ channel (GIRK1).

The Journal of Biological Chemistry
L H PhilipsonR J Miller

Abstract

An epitope-tagged form of an inwardly rectifying and G protein-coupled K+ channel (GIRK1-cp) was expressed at high levels in transfected mammalian cells. Immunoblot analysis of transfected human embryonic kidney cells (HEK293) and mouse insulinoma cells (beta TC3) revealed several GIRK1-cp polypeptides, including the major 59-kDa band, corresponding to the predicted mass of the GIRK1 polypeptide plus the epitope tag. Immunohistochemical staining using two anti-tag antibodies showed abundant immunoreactive material, which was predominantly concentrated in the perinuclear area in both transfected cell types. While functional GIRK1-cp message was present in poly(A)+ RNA prepared from HEK293 cells expressing GIRK1-cp protein, appropriate K+ currents could not be detected. In contrast, whole cell recordings made directly from transfected beta TC3 cells expressing GIRK1-cp revealed inwardly rectifying, pertussis toxin-sensitive currents activated by norepinephrine and galanin. Single channel recordings in excised patches of beta TC3 cells expressing GIRK1-cp showed rectifying K+ currents when activated by 50 microM guanosine 5'-O-(thiotriphosphate), with a slope conductance of 39.1 +/- 1.0 picosiemens. This is the first report of sta...Continue Reading

References

Oct 1, 1992·Trends in Biochemical Sciences·J R Hepler, A G Gilman
May 1, 1992·Archives of Biochemistry and Biophysics·K KontaniT Katada
Jan 1, 1990·Annual Review of Physiology·A M Brown, L Birnbaumer
Aug 1, 1990·Mayo Clinic Proceedings·D L LewisD E Clapham
Jul 1, 1991·British Journal of Pharmacology·K NakayamaM Hiraoka
Aug 1, 1991·Journal of Bioenergetics and Biomembranes·A M BrownL Birnbaumer
Jan 1, 1991·Proceedings of the National Academy of Sciences of the United States of America·L H PhilipsonD F Steiner
Jan 1, 1990·Cellular Signalling·S J Ashcroft, F M Ashcroft
Apr 1, 1989·Proceedings of the National Academy of Sciences of the United States of America·J R de WeilleM Lazdunski
Jun 26, 1989·Nucleic Acids Research·J Yon, M Fried
Dec 1, 1988·Proceedings of the National Academy of Sciences of the United States of America·S EfratS Baekkeskov
Aug 1, 1981·Pflügers Archiv : European journal of physiology·O P HamillF J Sigworth
Jul 1, 1994·The Journal of Cell Biology·W J ChangR G Anderson
Sep 1, 1994·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·R R Neubig
Nov 1, 1993·Proceedings of the National Academy of Sciences of the United States of America·N DascalD Trollinger
Jul 9, 1993·Science·R TaussigA G Gilman

❮ Previous
Next ❯

Citations

Jun 1, 1997·Brain Research. Molecular Brain Research·C S NelsonC N Allen
Jan 16, 1998·Cellular Signalling·N Dascal
Feb 3, 1999·Annals of the New York Academy of Sciences·T BranchekM W Walker
Jun 24, 1997·Proceedings of the National Academy of Sciences of the United States of America·M U EhrengruberN Davidson
Jan 1, 1997·Molecular and Cellular Neurosciences·E WischmeyerA Karschin
Sep 13, 2007·Journal of Cellular Physiology·Robert A Maue
Jan 21, 2010·Physiological Reviews·Hiroshi HibinoYoshihisa Kurachi
May 16, 2015·Journal of Cardiovascular Pharmacology·Said El-HaouJames T Milnes

❮ Previous
Next ❯

Related Concepts

Related Feeds

Adenoma, Islet Cell

Islet Cell Adenoma arises in the islet cells, which are insulin producing cells of the pancreas. These tumors can be either malignant or benign. Discover the latest research on Islet Cell Adenoma here.

ASBMB Publications

The American Society for Biochemistry and Molecular Biology (ASBMB) includes the Journal of Biological Chemistry, Molecular & Cellular Proteomics, and the Journal of Lipid Research. Discover the latest research from ASBMB here.