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1 Division of Nephrology, Hypertension and Endocrinology, Department of Medicine, Tohoku University Graduate School of Medicine, Sendai, Miyagi, Japan
2 Division of Nephrology, Hypertension and Endocrinology, Department of Medicine, Tohoku University Graduate School of Medicine, Sendai, Miyagi, Japan; PRESTO, Japan Science and Technology Agency, Kawaguchi, Saitama, Japan
3 Division of Gastroenterological Surgery, Department of Surgery, Tohoku University Graduate School of Medicine, Sendai, Miyagi, Japan
* To whom correspondence should be addressed. E-mail: mtanemoto-tky{at}umin.ac.jp.
Kir5.1, a non-functional inward-rectifying K+ channel by itself, can form functional channels by assembling with other proteins. We have previously shown that Kir5.1 assembled with Kir4.1 and functioned as an acid-base regulator in the kidney. In this study, we examined the intra-renal distribution of Kir5.1 by RT-PCR analysis on dissected nephron segments and immunohistochemical analysis with the specific anti-Kir5.1 antibody. Strong expression of Kir5.1 was detected in distal convoluted tubules, and weak expression was also detected in thick ascending limb of Henle's loop. Colocalization of Kir5.1 with Kir4.1 indicated expression of Kir5.1/Kir4.1 heteromer in these nephron segments. In a renal epithelial cell-line, MDCK cell, heteromer formation with Kir4.1 changed the localization of Kir5.1 from intracellular components to cell surface. The carboxyl-terminal cytoplasmic portion that includes PDZ binding motif of Kir4.1 was responsible for this intracellular localization. These data suggest the signals on the carboxyl-terminus of Kir4.1, including PDZ binding motif, determine the intracellular localization of Kir5.1/Kir4.1 heteromer in distal tubules.
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