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J. Biol. Chem., Vol. 281, Issue 38, 28254-28264, September 22, 2006

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Interaction of STIM1 with Endogenously Expressed Human Canonical TRP1 upon Depletion of Intracellular Ca²⁺ Stores^*

From the Department of Physiology (Cellular Physiology Research Group), University of Extremadura, 10071 Cáceres, Spain

STIM1 (stromal interaction molecule 1) has recently been proposed to communicate the intracellular Ca²⁺ stores with the plasma membrane to mediate store-operated Ca²⁺ entry. Here we describe for the first time that Ca²⁺ store depletion stimulates rapid STIM1 surface expression and association with endogenously expressed human canonical TRP1 (hTRPC1) independently of rises in cytosolic free Ca²⁺ concentration. These events require the support of the actin cytoskeleton in human platelets, as reported for the coupling between type II inositol 1,4,5-trisphosphate receptor in the Ca²⁺ stores and hTRPC1 in the plasma membrane, which has been suggested to underlie the activation of store-operated Ca²⁺ entry in these cells. Electrotransjection of cells with anti-STIM1 antibody, directed toward the N-terminal sequence that includes the Ca²⁺-binding region, prevented the migration of STIM1 toward the plasma membrane, the interaction between STIM1 and hTRPC1, the coupling between hTRPC1 and type II inositol 1,4,5-trisphosphate receptor, and reduced store-operated Ca²⁺ entry. These findings provide evidence for a role of STIM1 in the activation of store-operated Ca²⁺ entry probably acting as a Ca²⁺ sensor.

Received for publication, May 4, 2006 , and in revised form, July 26, 2006.

^* This work was supported by Ministerio de Educación y Ciencia-Dirección General de Investigación (MEC-DGI) Grant BFU2004-00165. 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.

¹ Supported by Dirección General de Investigación (DGI) Fellowship BES-2005-6899.

² To whom correspondence should be addressed: Dept. of Physiology, University of Extremadura, Cáceres 10071, Spain. Tel.: 34-927257154; Fax: 34-927257110; E-mail: jarosado{at}unex.es.

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