High-level expression and functional reconstitution of Shaker K+ channels

Biochemistry
T SunC Miller

Abstract

Voltage-gated K+ channels were expressed in COS cells transiently transfected with a plasmid carrying a cDNA for an inactivation-removed Shaker K+ channel driven by an adenovirus promoter. Channel expression was followed by immunological detection, binding of radioactive charybdotoxin (CTX), and functional reconstitution into planar lipid bilayers. About 10(7) channels per transfected cell are expressed on the plasma membrane. The expressed channels are glycosylated and competent to bind CTX with the expected characteristics. Channels observed after insertion into planar lipid bilayers displayed the voltage-dependent gating, conduction, and ion selectivity behavior expected for this channel. Channels were solubilized in several detergents without loss of CTX binding activity. The results make plausible a systematic attack on the purification of milligram-level amounts of functional K+ channels from a heterologous expression system.

References

Mar 15, 1991·Proceedings of the National Academy of Sciences of the United States of America·C S ParkC Miller
May 24, 1991·Science·C Miller
Mar 1, 1988·The Journal of General Physiology·R MacKinnon, C Miller
Oct 1, 1989·Proceedings of the National Academy of Sciences of the United States of America·R J LeonardH A Lester
Dec 1, 1987·Proceedings of the National Academy of Sciences of the United States of America·D D OprianH G Khorana
Jan 1, 1985·Annual Review of Biochemistry·R Kornfeld, S Kornfeld
Oct 1, 1993·Biophysical Journal·S A Goldstein, C Miller
Apr 1, 1994·Protein Engineering·E ShimonyC Miller
Feb 1, 1994·Quarterly Reviews of Biophysics·F J Sigworth
Feb 1, 1994·Current Biology : CB·M LiL Y Jan
Mar 1, 1994·Proceedings of the National Academy of Sciences of the United States of America·V E ScottJ O Dolly

❮ Previous
Next ❯

Citations

May 1, 2007·The Journal of Membrane Biology·Robert J O'ConnellSteven N Treistman
Oct 1, 2003·Journal of Molecular Biology·David N Parcej, Luise Eckhardt-Strelau
May 30, 2001·Progress in Biophysics and Molecular Biology·D Fedida, J C Hesketh
Aug 1, 1995·Quarterly Reviews of Biophysics·R Grisshammer, C G Tate
Mar 24, 2011·Nature Communications·Hui ZhengQiu-Xing Jiang
Apr 1, 1997·Proceedings of the National Academy of Sciences of the United States of America·M HannerM L Garcia
Mar 27, 1997·Biochemical and Biophysical Research Communications·R H Scannevin, J S Trimmer
Jun 11, 2004·Biophysical Journal·Chunbo YuanSteven N Treistman
May 3, 2000·Medical Hypotheses·A MansillaR Argüero
Nov 19, 2003·The Journal of Biological Chemistry·Leo A KimSteve A N Goldstein
Dec 1, 2006·Nature·Daniel SchmidtRoderick MacKinnon
Dec 26, 2001·The Journal of Biological Chemistry·U Subrahmanyeswara RaoPremalatha Balachandran
Jan 24, 1997·The Journal of Biological Chemistry·R H SpencerK G Chandy

❮ Previous
Next ❯

Related Concepts

Related Feeds

Cardiac Conduction System

The cardiac conduction system is a specialized tract of myocardial cells responsible for maintaining normal cardiac rhythm. Discover the latest research on the cardiac conduction system here.