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Am J Physiol Cell Physiol 280: C192-C198, 2001;
0363-6143/01 $5.00
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Vol. 280, Issue 1, C192-C198, January 2001

Differential renal distribution of NHERF isoforms and their colocalization with NHE3, ezrin, and ROMK

James B. Wade1, Paul A. Welling1, Mark Donowitz2, Shirish Shenolikar3, and Edward J. Weinman1,4,5

Departments of 1 Physiology and 4 Medicine, Division of Nephrology, University of Maryland School of Medicine and 5 Medical Service, Department of Veterans Affairs Medical Center, Baltimore 21201; 2 Gastroenterology Division, Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205; and 3 Department of Pharmacology and Cancer Biology, Duke University School of Medicine, Durham, North Carolina 27710

Na+/H+ exchanger regulatory factor (NHERF) and NHERF2 are PDZ motif proteins that mediate the inhibitory effect of cAMP on Na+/H+ exchanger 3 (NHE3) by facilitating the formation of a multiprotein signaling complex. With the use of antibodies specific for NHERF and NHERF2, immunocytochemical analysis of rat kidney was undertaken to determine the nephron distribution of both proteins and their colocalization with other transporters and with ezrin. NHERF was most abundant in apical membrane of proximal tubule cells, where it colocalized with ezrin and NHE3. NHERF2 was detected in the glomerulus and in other renal vascular structures. In addition, NHERF2 was strongly expressed in collecting duct principal cells, where it colocalized with ROMK. These results indicate a striking difference in the nephron distribution of NHERF and NHERF2 and suggests NHERF is most likely to be the relevant biological regulator of NHE3 in the proximal tubule, while NHERF2 may interact with ROMK or other targets in the collecting duct. The finding that NHERF isoforms occur in different cell types suggests that NHERF and NHERF2 may subserve different functions in the kidney.

sodium hydrogen exchanger regulatory protein 2; sodium hydrogen exchanger 3 protein kinase A regulatory protein; signal complexes; PDZ proteins


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