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J. Biol. Chem., Vol. 279, Issue 45, 46962-46968, November 5, 2004

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PSD-95 and Lin-7b Interact with Acid-sensing Ion Channel-3 and Have Opposite Effects on H⁺-gated Current^*

Alesia M. Hruska-Hageman¶, Christopher J. Benson, A. Soren Leonard, Margaret P. Price, and Michael J. Welsh, An investigator of the Howard Hughes Medical Institute||**

From the Departments of Internal Medicine and ^||Physiology and Biophysics and the Howard Hughes Medical Institute, Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, Iowa 52242

The acid-sensing ion channel-3 (ASIC3) is a degenerin/epithelial sodium channel expressed in the peripheral nervous system. Previous studies indicate that it participates in the response to mechanical and painful stimuli, perhaps contributing to mechanoreceptor and/or H⁺-gated nociceptor function. ASIC3 subunits contain intracellular N and C termini that may control channel localization and function. We found that a PDZ-binding motif at the ASIC3 C terminus interacts with four different proteins that contain PDZ domains: PSD-95, Lin-7b, MAGI-1b, and PIST. ASIC3 and these interacting proteins were expressed in dorsal root ganglia and spinal cord, and PSD-95 co-precipitated ASIC3 from spinal cord. When expressed in heterologous cells, PSD-95 reduced the amplitude of ASIC3 acid-evoked currents, whereas Lin-7b increased current amplitude. PSD-95 and Lin-7b altered current density by decreasing or increasing, respectively, the amount of ASIC3 on the cell surface. The finding that multiple PDZ-containing proteins bind ASIC3 and can influence its presence in the plasma membrane suggests that they may play an important role in the contribution of ASIC3 to nociception and mechanosensation.

Received for publication, May 26, 2004 , and in revised form, August 13, 2004.

^* Work done in the In Vitro Models and Cell Culture Core was supported by the NHLBI, National Institutes of Health; by Cystic Fibrosis Foundation Grants R458-CR02 and ENGLH9850; and by NIDDK, National Institutes of Health Grant DK54759. 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.

^¶ An associate of the Howard Hughes Medical Institute.

^** To whom correspondence should be addressed: Howard Hughes Medical Inst., Roy J. and Lucille A. Carver College of Medicine, 500 EMRB, University of Iowa, Iowa City, IA 52242. Tel.: 319-335-7619; Fax: 319-335-7623; E-mail: michael-welsh{at}uiowa.edu.

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