A photoactivatable probe for the Na+/H+ exchanger cross-links a 66-kDa renal brush border membrane protein

W. Ross, W. Bertrand, A. Morrison

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14 Scopus citations

Abstract

Earlier studies on LLC-PK1 cells have demonstrated two pharmacologically distinct Na+/H+ exchangers in renal epithelia. In addition, the cDNA clone for the human Na+/H+ antiporter which is growth factor activatable has been isolated and expressed (Sardet, C., Franchi, A., and Pouyssegur, J. (1989) Cell 56, 271-280). We report here the synthesis of an amiloride analogue that can be photoactivated and labeled with 125I. This analogue covalently cross-links a 66-kDa protein of bovine brush border membranes. A rabbit polyclonal antibody that was directed against a 20-amino acid peptide of the cytoplasmic domain of its human Na+/H+ antiporter also gives a positive Western against 66-kDa protein of bovine brush border membranes. Thus, the photoactive probe may be helpful in the isolation and purification of the brush border Na+/H+ exchanger.

Original languageEnglish
Pages (from-to)5341-5344
Number of pages4
JournalJournal of Biological Chemistry
Volume265
Issue number10
StatePublished - 1990

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