The Journal of General Physiology
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Published online Jan 30 2006. doi:10.1085/jgp.200509370
The Rockefeller University Press, 0022-1295 $8.00
JGP, Volume 127, Number 2, 191-204
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ARTICLE

Structural Determinants for Functional Coupling Between the ß and {alpha} Subunits in the Ca2+-activated K+ (BK) Channel

Patricio Orio1, Yolima Torres1,2, Patricio Rojas1,3, Ingrid Carvacho1,4, Maria L. Garcia5, Ligia Toro6, Miguel A. Valverde7, and Ramon Latorre1,3

1 Department of Biophysics and Molecular Physiology, Centro de Estudios Científicos, Valdivia 5110246, Chile
2 Universidad del Valle, Cali, Colombia
3 Facultad de Ciencias, Universidad de Chile, Santiago 7800024, Chile
4 Universidad Austral de Chile, Valdivia, Chile
5 Department of Ion Channels, Merck Research Laboratories, Rahway, NJ 07065
6 Department of Anesthesiology University of California, Los Angeles, CA 90095
7 Grup de Canalopaties, Unitat de Senyalització Celular, Universitat Pompeu Fabra, Barcelona 08003, Spain

High conductance, calcium- and voltage-activated potassium (BK, MaxiK) channels are widely expressed in mammals. In some tissues, the biophysical properties of BK channels are highly affected by coexpression of regulatory (ß) subunits. The most remarkable effects of ß1 and ß2 subunits are an increase of the calcium sensitivity and the slow down of channel kinetics. However, the detailed characteristics of channels formed by {alpha} and ß1 or ß2 are dissimilar, the most remarkable difference being a reduction of the voltage sensitivity in the presence of ß1 but not ß2. Here we reveal the molecular regions in these ß subunits that determine their differential functional coupling with the pore-forming {alpha}-subunit. We made chimeric constructs between ß1 and ß2 subunits, and BK channels formed by {alpha} and chimeric ß subunits were expressed in Xenopus laevis oocytes. The electrophysiological characteristics of the resulting channels were determined using the patch clamp technique. Chimeric exchange of the different regions of the ß1 and ß2 subunits demonstrates that the NH3 and COOH termini are the most relevant regions in defining the behavior of either subunit. This strongly suggests that the intracellular domains are crucial for the fine tuning of the effects of these ß subunits. Moreover, the intracellular domains of ß1 are responsible for the reduction of the BK channel voltage dependence. This agrees with previous studies that suggested the intracellular regions of the {alpha}-subunit to be the target of the modulation by the ß1-subunit.


P. Orio and Y. Torres contributed equally to this work.

P. Orio's present address is the Instituto de Neurociencias de Alicante, Universidad Miguel Hernandez-CSIC, 03550 Sant Joan d'Alacant, Spain.

P. Rojas's present address is the Department of Anatomy and Neurobiology, Washington University School of Medicine, St. Louis, MO 63110


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