title: Allosteric modulation by ATP of the bovine adrenal natriuretic factor R1 receptor functions.
abstract: Atrial natriuretic factor (ANF-R1) receptor is a 130-kDa protein that contains a cytoplasmic guanylate cyclase domain. We report that ATP interacts in an allosteric manner with the ANF-R1 receptor, resulting in reduced ANF binding and enhanced ANF-stimulated guanylate cyclase activity. The modulatory properties of various nucleotides indicate a preference for the adenine family with a rank order of potency of ATP greater than App(NH)p greater than or equal to ADP greater than or equal to AMP while cyclic and guanine nucleotides except GTP are inactive. The negative modulation by ATP of ANF binding is specific for the ANF-R1 receptor subtype since the amount of ANF bound by the guanylate cyclase uncoupled ANF-R2 subtype is increased in the presence of ATP. Furthermore, the effects of ATP on ANF-R1 receptor binding function are still observed with the affinity-purified ANF-R1 receptor, suggesting an allosteric binding site for ATP on the ANF-R1 receptor. In intact membranes, limited proteolysis of the ANF-R1 receptor with trypsin dose-dependently prevents the ATP-induced decrease in ANF binding concomitantly with the formation of a membrane-associated ANF-binding fragment of 70 kDa. These results confirm the direct modulatory role of ATP on hormone binding activity of ANF-R1 receptor and suggest that the nucleotide regulatory binding site is located in the intracellular domain vicinal to the protease-sensitive region.
authors.affiliation: Department of Pharmacology, University of Montreal, Quebec, Canada.
authors.forename: L
authors.initials: L
authors.lastname: Larose
authors.forename: N
authors.initials: N
authors.lastname: McNicoll
authors.forename: H
authors.initials: H
authors.lastname: Ong
authors.forename: A
authors.initials: A
authors.lastname: De Léan
mesh_terms: D000255:Adenosine Triphosphate / Q000494:pharmacology*; D000495:Allosteric Site; D000818:Animals; D009320:Atrial Natriuretic Factor / Q000378:metabolism*; D002417:Cattle; D003593:Cytoplasm / Q000378:metabolism; D006868:Hydrolysis; D017461:Receptors, Atrial Natriuretic Factor; D011956:Receptors, Cell Surface / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D014357:Trypsin / Q000494:pharmacology; D015384:Zona Glomerulosa / Q000187:drug effects / Q000378:metabolism*
chemical_list: D011956:Receptors, Cell Surface; D009320:Atrial Natriuretic Factor; D000255:Adenosine Triphosphate; D014357:Trypsin; D017461:Receptors, Atrial Natriuretic Factor