title: Human kidney amiloride-binding protein: cDNA structure and functional expression.
abstract: 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.
authors.affiliation: Institut de Pharmacologie Moléculaire et Cellulaire, Centre National de la Recherche Scientifique, Valbonne, France.
authors.forename: P
authors.initials: P
authors.lastname: Barbry
authors.forename: M
authors.initials: M
authors.lastname: Champe
authors.forename: O
authors.initials: O
authors.lastname: Chassande
authors.forename: S
authors.initials: S
authors.lastname: Munemitsu
authors.forename: G
authors.initials: G
authors.lastname: Champigny
authors.forename: E
authors.initials: E
authors.lastname: Lingueglia
authors.forename: P
authors.initials: P
authors.lastname: Maes
authors.forename: C
authors.initials: C
authors.lastname: Frelin
authors.forename: A
authors.initials: A
authors.lastname: Tartar
authors.forename: A
authors.initials: A
authors.lastname: Ullrich
mesh_terms: D000584:Amiloride / Q000031:analogs & derivatives / Q000378:metabolism; D006631:Amine Oxidase (Copper-Containing); D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002352:Carrier Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics*; D015723:Gene Library; D006801:Humans; D007700:Kinetics; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D010449:Peptide Mapping; D012333:RNA, Messenger / Q000235:genetics / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000378:metabolism; D012689:Sequence Homology, Nucleic Acid; D013552:Swine; D014162:Transfection
chemical_list: D002352:Carrier Proteins; D015345:Oligonucleotide Probes; D012333:RNA, Messenger; D011994:Recombinant Proteins; C047088:phenylamil; D000584:Amiloride; D004247:DNA; D006631:Amine Oxidase (Copper-Containing); C065976:AOC1 protein, human