Human kidney amiloride-binding protein: cDNA structure and functional expression

Journal Article · Mon Oct 01 00:00:00 EDT 1990 · Proceedings of the National Academy of Sciences of the United States of America; (USA)

DOI:https://doi.org/10.1073/pnas.87.19.7347· OSTI ID:5803801

Barbry, P; Chassande, O; Champigny, G; Lingueglia, E; Frelin, C; Lazdunski, M ^[1]; Champe, M; Munemitsu, S; Ullrich, A ^[2]; Maes, P; Tartar, A ^[3]

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1. Centre National de la Recherche Scientifique, Valbonne (France)
2. Genentech, Inc., San Francisco, CA (USA)
3. Centre National de la Recherche Scientifique, Lille (France) Institut Pasteur de Lille (France)

Phenamil, an analog of amiloride, is a potent blocker of the epithelial Na{sup plus} 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{sup plus} channel in pig kidney membranes and (ii) that is immunoprecipitated with monoclonal antibodies raised against pig kidney amiloride-binding protein.

• OSTI ID: 5803801
• Journal Information: Proceedings of the National Academy of Sciences of the United States of America; (USA), Vol. 87:19; ISSN 0027-8424
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
CATIONS
MEMBRANE TRANSPORT
MEMBRANE PROTEINS
AMINO ACID SEQUENCE
RECOMBINANT DNA
DNA SEQUENCING
DIURETICS
DNA-CLONING
KIDNEYS
MESSENGER-RNA
MONOCLONAL ANTIBODIES
OLIGONUCLEOTIDES
SODIUM COMPOUNDS
SULFUR 35
TRITIUM COMPOUNDS
ALKALI METAL COMPOUNDS
ANTIBODIES
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BODY
CHARGED PARTICLES
CLONING
DAYS LIVING RADIOISOTOPES
DNA
DNA HYBRIDIZATION
DRUGS
EVEN-ODD NUCLEI
HYBRIDIZATION
HYDROGEN COMPOUNDS
IONS
ISOTOPES
LIGHT NUCLEI
MOLECULAR STRUCTURE
NUCLEI
NUCLEIC ACIDS
ORGANIC COMPOUNDS
ORGANS
PROTEINS
RADIOISOTOPES
RNA
STRUCTURAL CHEMICAL ANALYSIS
SULFUR ISOTOPES
550201* - Biochemistry- Tracer Techniques