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J Biol Chem, Vol. 274, Issue 7, 4383-4388, February 12, 1999

Molecular Cloning and Functional Expression of a Rat Na+/H+ Exchanger (NHE5) Highly Expressed in Brain

Surat AttaphitayaDagger §, Keerang ParkDagger , and James E. MelvinDagger

From the Dagger  Center for Oral Biology and the § Department of Neurobiology and Anatomy, University of Rochester School of Medicine and Dentistry, Rochester, New York 14642

We report here the cloning, primary structure, heterologous expression, tissue distribution, and localization of a cDNA encoding rat NHE5, a fifth member of the mammalian plasma membrane Na+/H+ exchanger (NHE) gene family. The full-length open reading frame as well as 34 nucleotides of 5'-untranslated and 1443 nucleotides of 3'-untranslated sequences were obtained using a polymerase chain reaction strategy involving reverse transcription-polymerase chain reaction and 5'/3'-rapid amplification of cDNA ends. The NHE5 cDNA encodes a protein of 898 amino acids with a calculated Mr of 99,044 and is predicted to contain 11-13 transmembrane domains. An amino acid comparison of the coding region of rat NHE5 reveals 95% identity with human NHE5. Northern hybridization analysis showed that high level expression of NHE5 mRNA is restricted to brain. Transfection of the coding region of rat NHE5 into NHE-deficient PS120 cells resulted in Na+/H+ exchange activity that was relatively insensitive to the amiloride analogue, 5-(N-ethyl-N-isopropyl) amiloride, with a half-maximal inhibitory concentration (IC50) of 1.53 ± 0.25 µM. In situ hybridization of rat brain sections revealed significant NHE5 mRNA levels in the dentate gyrus with lower levels observed in the hippocampus and cerebral cortex. These results suggest a specialized role for this fifth NHE isoform in neuronal tissues.

Copyright © 1999 by The American Society for Biochemistry and Molecular Biology, Inc.
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