PDZ binding motif-dependent localization of K+ channel on the basolateral side in distal tubules.
Kir5.1, a nonfunctional inwardly rectifying K(+) channel by itself, can form functional channels by assembling with other proteins. We previously showed that Kir5.1 assembled with Kir4.1 and functioned as an acid-base regulator in the kidney. In this study, we examined the intrarenal 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, Madin-Darby canine kidney cells, heteromer formation with Kir4.1 changed the localization of Kir5.1 from intracellular components to the cell surface. The COOH-terminal cytoplasmic portion that includes the PDZ binding motif of Kir4.1 was responsible for this intracellular localization. These data suggest the signals on the COOH terminus of Kir4.1, including PDZ binding motif, determine the intracellular localization of Kir5.1/Kir4.1 heteromer in distal tubules.
Tanemoto
Masayuki
M
Division of Nephrology, Hypertension and Endocrinology, Department of Medicine, Tohoku University Graduate School of Medicine, 1-1 Seiryo-cho, Aoba-ku, Sendai 980-8574, Japan. mtanemoto-tky@umin.ac.jp
Abe
Takaaki
T
Onogawa
Tohru
T
Ito
Sadayoshi
S
D000818:Animals; D000918:Antibody Specificity; D001665:Binding Sites; D002460:Cell Line; D004285:Dogs; D005455:Fluorescent Antibody Technique; D007150:Immunohistochemistry; D007668:Kidney / Q000737:chemistry; D007686:Kidney Tubules, Distal / Q000737:chemistry*; D008297:Male; D018613:Microscopy, Confocal; D009399:Nephrons / Q000737:chemistry; D024661:Potassium Channels, Inwardly Rectifying / Q000032:analysis* / Q000737:chemistry* / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014018:Tissue Distribution; D000097062:Kir5.1 Channel; D000098854:Kcnj10 Channel
D024661:Potassium Channels, Inwardly Rectifying; D012333:RNA, Messenger; D000097062:Kir5.1 Channel; D000098854:Kcnj10 Channel