Am J Physiol Cell Physiol AJP: Endocrinology and Metabolism
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Am J Physiol Cell Physiol 281: C2039-C2048, 2001;
0363-6143/01 $5.00
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Vol. 281, Issue 6, C2039-C2048, December 2001

Half-lives of plasma membrane Na+/H+ exchangers NHE1-3: plasma membrane NHE2 has a rapid rate of degradation

Megan E. Cavet, Shafinaz Akhter, Rakhilya Murtazina, Fermin Sanchez de Medina, Chung-Ming Tse, and Mark Donowitz

Departments of Medicine and Physiology, Gastrointestinal Division, The Johns Hopkins University School of Medicine, Baltimore, Maryland 21205

The Na+/H+ exchangers NHE2 and NHE3 are involved in epithelial Na+ and HCO<UP><SUB>3</SUB><SUP>−</SUP></UP> absorption. To increase insights into the functions of NHE2 vs. NHE3, we compared their cellular processing with each other and with the housekeeping isoform NHE1. Using biotinylated exchanger, we determined that the half-life of plasma membrane NHE2 was short (3 h) compared with that of NHE1 (24 h) and NHE3 (14 h) in both PS120 fibroblasts and Caco-2 cells. NHE2 transport and plasma membrane levels were reduced by 3 h of Brefeldin A treatment, whereas NHE1 was unaffected. NHE2 was degraded by the lysosomes but not proteosomes, as demonstrated by increasing levels of endocytosed NHE2 protein after inhibition of the lysosomes, but not with proteosome inhibition. Unlike that of NHE3, basal NHE2 transport activity was not affected by phosphatidylinositol 3-kinase inhibition and did not appear to be localized in the juxtanuclear recycling endosome. Therefore, for NHE2, protein degradation and/or protein synthesis probably play important roles in its basal and regulated states. These results suggest fundamental differences in the cellular processing and trafficking of NHE2 and NHE3. These differences may underlie the specialized roles that these exchangers play in epithelial cells.

sodium absorption; Caco-2 cells; lysosomes; trafficking


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