Human Kidney Amiloride-Binding Protein: cDNA Structure and Functional Expression

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 87; no. 19; pp. 7347 - 7351
• Main Authors: Barbry, Pascal, Champe, Mark, Chassande, Olivier, Munemitsu, Susan, Champigny, Guy, Lingueglia, Eric, Maes, Pierrette, Frelin, Christian, Tartar, Andre, Ullrich, Axel, Lazdunski, Michael
• Format: Journal Article
• Language: English
• Published: Washington, DC National Academy of Sciences of the United States of America 01-10-1990; National Acad Sciences
• Subjects: 3T3 cells; 550201 - Biochemistry- Tracer Techniques; ALKALI METAL COMPOUNDS; Amiloride - analogs & derivatives; Amiloride - metabolism; Amine Oxidase (Copper-Containing); AMINO ACID SEQUENCE; Amino acids; Animals; ANTIBODIES; Base Sequence; BASIC BIOLOGICAL SCIENCES; BETA DECAY RADIOISOTOPES; BETA-MINUS DECAY RADIOISOTOPES; Biochemistry; Biological and medical sciences; BODY; Carrier Proteins - genetics; Carrier Proteins - isolation & purification; Carrier Proteins - metabolism; CATIONS; Cell Line; Cell lines; Cell Membrane - metabolism; CHARGED PARTICLES; CLONING; Cloning, Molecular; Complementary DNA; DAYS LIVING RADIOISOTOPES; DIURETICS; DNA; DNA - genetics; DNA HYBRIDIZATION; DNA SEQUENCING; DNA-CLONING; DRUGS; EVEN-ODD NUCLEI; Fundamental and applied biological sciences. Psychology; Gene Library; genes; Humans; HYBRIDIZATION; HYDROGEN COMPOUNDS; IONS; ISOTOPES; Kidney cells; KIDNEYS; Kinetics; LIGHT NUCLEI; MEMBRANE PROTEINS; MEMBRANE TRANSPORT; MESSENGER-RNA; Molecular and cellular biology; Molecular genetics; Molecular Sequence Data; MOLECULAR STRUCTURE; MONOCLONAL ANTIBODIES; NIH 3T3 cells; NUCLEI; NUCLEIC ACIDS; Oligonucleotide Probes; OLIGONUCLEOTIDES; ORGANIC COMPOUNDS; ORGANS; Peptide Mapping; PROTEINS; RADIOISOTOPES; Receptors; RECOMBINANT DNA; Recombinant Proteins - metabolism; RNA; RNA, Messenger - genetics; RNA, Messenger - isolation & purification; Sequence Homology, Nucleic Acid; SODIUM COMPOUNDS; STRUCTURAL CHEMICAL ANALYSIS; SULFUR 35; SULFUR ISOTOPES; Swine; Transfection; TRITIUM COMPOUNDS; Human; Binding protein; Primary structure; Complementary DNA; Kidney
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.87.19.7347

Phenamil, an analog of amiloride, is a potent blocker of the epithelial Na+channel. It has been used to purify the porcine kidney amiloride-binding protein. Synthetic oligonucleotides derived from partial sequences have been used to screen a human kidney cDNA library and to isolate the cDNA encoding the human amiloride-binding protein. The primary structure was deduced from the DNA sequence analysis. The protein is 713 residues long, with a 19-amino acid signal peptide. The mRNA was expressed in 293-S and NIH 3T3 cells, yielding a glycoprotein (i) that binds amiloride and amiloride analogs with affinities similar to the amiloride receptor associated with the apical Na+channel in pig kidney membranes and (ii) that is immunoprecipitated with monoclonal antibodies raised against pig kidney amiloride-binding protein.