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

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. 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 renal 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.
Authors:
; ;  [1]
  1. (Washington Univ. School of Medicine, St. Louis, MO (USA))
Publication Date:
OSTI Identifier:
7118073
Resource Type:
Journal Article
Resource Relation:
Journal Name: Journal of Biological Chemistry; (USA); Journal Volume: 265:10
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; DIURETICS; BIOLOGICAL FUNCTIONS; MEMBRANE PROTEINS; CROSS-LINKING; ANTIBODIES; CATTLE; CHEMICAL PREPARATION; DOGS; IODINE 125; KIDNEYS; MASS SPECTRA; MOLECULAR WEIGHT; PROTEIN STRUCTURE; RABBITS; REAGENTS; ANIMALS; BETA DECAY RADIOISOTOPES; BODY; CHEMICAL REACTIONS; DAYS LIVING RADIOISOTOPES; DOMESTIC ANIMALS; DRUGS; ELECTRON CAPTURE RADIOISOTOPES; FUNCTIONS; INTERMEDIATE MASS NUCLEI; IODINE ISOTOPES; ISOTOPES; MAMMALS; NUCLEI; ODD-EVEN NUCLEI; ORGANIC COMPOUNDS; ORGANS; POLYMERIZATION; PROTEINS; RADIOISOTOPES; RUMINANTS; SPECTRA; SYNTHESIS; VERTEBRATES 550201* -- Biochemistry-- Tracer Techniques