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J. Biol. Chem., Vol. 279, Issue 28, 29263-29269, July 9, 2004

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Plasma Membrane Expression of T-type Calcium Channel ₁ Subunits Is Modulated by High Voltage-activated Auxiliary Subunits^*

Stefan J. Dubel, Christophe Altier, Séverine Chaumont, Philippe Lory, Emmanuel Bourinet, and Joël Nargeot¶

From the Département de Physiologie, Laboratoire de Génomique Fonctionnelle, CNRS-Unité Propre de Recherche 2580, 34396 Montpellier, France

It has been suggested that the auxiliary subunits of high voltage-activated (HVA) calcium channels modulate T-type, low voltage-activated (LVA) calcium channels. Such a regulation has yet to be documented, especially because there has been no biochemical characterization of T-channels. To monitor total protein levels and plasma membrane expression of T-channels in living cells, external epitopes (hemagglutinin, FLAG) were introduced into human recombinant Ca_V3 channels that were also N-terminally fused to green fluorescent protein. Utilizing Western blot techniques, fluorescence flow cytometry, immunofluorescence, luminometry, and electrophysiology, we describe here that _1b and ₂-₁ subunits enhance the level of Ca_V3 proteins as well as their plasma membrane expression in various expression systems. We also report that, in both Xenopus oocytes and mammalian cells, the ₂-₁ subunits increase by at least and _1b 2-fold the current density of Ca_V3 channels with no change in the electrophysiological properties. Altogether, these data indicate that HVA auxiliary subunits modulate Ca_V3 channel surface expression, suggesting that the membrane targeting of HVA and LVA ₁ subunits is regulated dynamically through the expression of a common set of regulatory subunits.

Received for publication, December 9, 2003 , and in revised form, April 7, 2004.

^* This work was supported in part by CNRS and grants from the Institut UPSA de la Douleur, the Paul Hamel foundation, the Association Française contre les Myopathies (AFM), and the Association pour la Recherche sur le Cancer (ARC). 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 fellowship from the Region Languedoc-Roussillon.

^¶ To whom correspondence should be addressed: Dépt. de Physiologie, Laboratoire de Génomique Fonctionnelle, CNRS-Unité Propre de Recherche 2580, 141 rue de la Cardonille, 34396 Montpellier Cedex 01, France. Tel.: 33-4-99-61-99-67; Fax: 33-4-99-61-99-01; E-mail: joel.nargeot{at}igh.cnrs.fr.

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