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J. Biol. Chem., Vol. 279, Issue 35, 36445-36453, August 27, 2004

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Coassembly of Big Conductance Ca²⁺-activated K⁺ Channels and L-type Voltage-gated Ca²⁺ Channels in Rat Brain^*

Morten Grunnet¶ and Walter A. Kaufmann||

From the Department of Medical Physiology, the Panum Institute, University of Copenhagen, Blegdamsvej 3, Copenhagen DK-2200, Denmark, Neurosearch A/S, Pederstrupsvej 93, Ballerup 2750, Denmark, and the ^||Department of Physiology, Centre for Molecular Biology and Neuroscience, University of Oslo, Oslo N-0317, Norway

Based on electrophysiological studies, Ca²⁺-activated K⁺ channels and voltage-gated Ca²⁺ channels appear to be located in close proximity in neurons. Such colocalization would ensure selective and rapid activation of K⁺ channels by local increases in the cytosolic calcium concentration. The nature of the apparent coupling is not known. In the present study we report a direct coassembly of big conductance Ca²⁺-activated K⁺ channels (BK) and L-type voltage-gated Ca²⁺ channels in rat brain. Saturation immunoprecipitation studies were performed on membranes labeled for BK channels and precipitated with antibodies against _1C and _1D L-type Ca²⁺ channels. To confirm the specificity of the interaction, precipitation experiments were carried out also in reverse order. Also, additive precipitation was performed because _1C and _1D L-type Ca²⁺ channels alrefer ways to separate ion channel complexes. Finally, immunochemical studies showed a distinct but overlapping expression pattern of the two types of ion channels investigated. BK and L-type Ca²⁺ channels were colocalized in various compartments throughout the rat brain. Taken together, these results demonstrate a direct coassembly of BK channels and L-type Ca²⁺ channels in certain areas of the brain.

Received for publication, March 1, 2004 , and in revised form, June 8, 2004.

^* The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

^¶ Supported by a traveling scholarship from the University of Copenhagen. To whom correspondence should be addressed: Dept. of Medical Physiology, 12.6.26, The Panum Institute, University of Copenhagen, Blegdamsvej 3, Copenhagen DK-2200, Denmark. Tel.: 45-3532-7565; Fax: 45-3532-7526; E-mail: mgrunnet{at}mfi.ku.dk.

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