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J. Biol. Chem., Vol. 282, Issue 14, 10585-10593, April 6, 2007

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Identification of a Pool of Non-pumping Na/K-ATPase^*

Man Liang¹, Jiang Tian¹, Lijun Liu, Sandrine Pierre, Jiang Liu, Joseph Shapiro, and Zi-Jian Xie²

From the Department of Physiology, Pharmacology, Metabolism, and Cardiovascular Sciences and the Department of Medicine, University of Toledo Health Science Campus, Toledo, Ohio 43614

Recent studies have ascribed many non-pumping functions to the Na/K-ATPase. Here, we present experimental evidence demonstrating that over half of the plasma membrane Na/K-ATPase in LLC-PK1 cells is performing cellular functions other than ion pumping. This "non-pumping" pool of Na/K-ATPase, like the pumping pump, binds ouabain. Depletion of either cholesterol or caveolin-1 moves some of the "non-pumping" Na/K-ATPase into the pumping pool. Graded knock-down of the 1 subunit of the Na/K-ATPase eventually results in loss of this "non-pumping" pool while preserving the pumping pool. Our prior studies indicate that a loss of the non-pumping pool is associated with a loss of receptor function as evidenced by the failure of ouabain administration to induce the activation of Src and/or ERK. Therefore, our new findings suggest that a substantial amount of surface-expressed Na/K-ATPase, at least in some types of cells, may function as non-canonical ouabain-binding receptors.

Received for publication, September 27, 2006 , and in revised form, February 6, 2007.

^* This work was supported in part by by National Institutes of Health Grants HL-36573 and HL-67963, awarded by NHLBI, United States Public Health Service, Dept. of Health and Human Services. 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 These authors contributed equally to this work.

² To whom correspondence should be addressed: Dept. of Physiology, Pharmacology, Metabolism, and Cardiovascular Sciences, The University of Toledo Health Science Campus, Mail Stop 1008, 3000 Arlington Ave., Toledo, OH 43614-2598. Tel.: 419-383-4182; Fax: 419-383-2871; E-mail: zxie{at}meduohio.edu.

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