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Originally published In Press as doi:10.1074/jbc.M106123200 on July 12, 2001

J. Biol. Chem., Vol. 276, Issue 39, 36673-36680, September 28, 2001
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Requirement of Multiple Protein Domains and Residues for Gating KATP Channels by Intracellular pH*

Hailan PiaoDagger , Ningren CuiDagger , Haoxing Xu, Jinzhe Mao, Asheebo Rojas, Runping Wang, Latifat Abdulkadir, Lilly Li, Jiangping Wu, and Chun Jiang§

From the Department of Biology, Georgia State University, Atlanta, Georgia 30302-4010

ATP-sensitive K+ channels (KATP) are regulated by pH in addition to ATP, ADP, and phospholipids. In the study we found evidence for the molecular basis of gating the cloned KATP by intracellular protons. Systematic constructions of chimerical Kir6.2-Kir1.1 channels indicated that full pH sensitivity required the N terminus, C terminus, and M2 region. Three amino acid residues were identified in these protein domains, which are Thr-71 in the N terminus, Cys-166 in the M2 region, and His-175 in the C terminus. Mutation of any of them to their counterpart residues in Kir1.1 was sufficient to completely eliminate the pH sensitivity. Creation of these residues rendered the mutant channels clear pH-dependent activation. Thus, critical players in gating KATP by protons are demonstrated. The pH sensitivity enables the KATP to regulate cell excitability in a number of physiological and pathophysiological conditions when pH is low but ATP concentration is normal.

* This work was supported by National Institutes of Health Grant HL58410, American Diabetes Association Grant 1-01-RA-12, and American Heart Association Grant 9950528N.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Dagger These authors contributed equally to this work.

§ To whom correspondence should be addressed: Dept. of Biology, Georgia State University, 24 Peachtree Center Ave., Atlanta, GA 30302-4010. Tel.: 404-651-0913; Fax: 404-651-2509; E-mail: cjiang@gsu.edu.

Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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