title: Purification and characterization of the 180-kDa membrane guanylate cyclase containing atrial natriuretic factor receptor from rat adrenal gland and its regulation by protein kinase C.
abstract: The original concept that cyclic GMP is one of the mediators of the hormone-dependent process of steroidogenesis has been strengthened by the characterization of a 180-kDa protein from rat adrenocortical carcinoma and rat and mouse testes. This protein appears to have an unusual characteristic of containing both the atrial natriuretic factor (ANF)-binding and guanylate cyclase activities, and appears to be intimately involved in the ANF-dependent steroidogenic signal transduction. In rat adrenal glands we now demonstrate: 1) the direct presence of a 180-kDa ANF-binding protein in GTP-affinity purified membrane fraction as evidenced by affinity cross-linking technique and by the Western blot analysis of the partially purified enzyme; 2) that the enzyme is biochemically and immunologically different from the soluble guanylate cyclase as there is no antigenic cross-reactivity of 180-kDa guanylate cyclase antibody with soluble guanylate cyclase; 3) in contrast to the soluble guanylate cyclase, the particulate enzyme is not stimulated by nitrite-generating compounds and hemin; and 4) protein kinase C inhibits both the basal and ANF-dependent guanylate cyclase activity and phosphorylates the 180-kDa guanylate cyclase. These results reveal the presence of a 180-kDa protein in rat adrenal glands and support the contention that: (a) this protein contains both the guanylate cyclase and ANF receptor; (b) the 180-kDa enzyme is coupled with the ANF-dependent cyclic GMP production; (c) the 180-kDa enzyme is biochemically distinct from the nonspecific soluble guanylate cyclase; and (d) there is a protein kinase C-dependent negative regulatory loop for the operation of ANF-dependent cyclic GMP signal pathway which acts via the phosphorylation of 180-kDa guanylate cyclase.
authors.affiliation: Section of Regulatory Biology, Cleveland Clinic Research Institute, OH 44195-5068.
authors.forename: R K
authors.initials: RK
authors.lastname: Sharma
authors.forename: R B
authors.initials: RB
authors.lastname: Marala
authors.forename: T M
authors.initials: TM
authors.lastname: Duda
mesh_terms: D000306:Adrenal Cortex Neoplasms / Q000276:immunology; D000311:Adrenal Glands / Q000378:metabolism / Q000648:ultrastructure*; D000818:Animals; D000906:Antibodies / Q000032:analysis; D009320:Atrial Natriuretic Factor / Q000378:metabolism; D001386:Azides / Q000494:pharmacology; D006152:Cyclic GMP / Q000096:biosynthesis; D006162:Guanylate Cyclase / Q000302:isolation & purification*; D006427:Hemin / Q000494:pharmacology; D006454:Hemoglobins / Q000494:pharmacology; D009599:Nitroprusside / Q000494:pharmacology; D010766:Phosphorylation; D011493:Protein Kinase C / Q000378:metabolism; D011817:Rabbits; D051381:Rats; D017461:Receptors, Atrial Natriuretic Factor; D011956:Receptors, Cell Surface / Q000032:analysis* / Q000378:metabolism; D019810:Sodium Azide
chemical_list: D000906:Antibodies; D001386:Azides; D006454:Hemoglobins; D011956:Receptors, Cell Surface; C472610:atrial natriuretic peptide, rat; D009599:Nitroprusside; D006427:Hemin; D009320:Atrial Natriuretic Factor; D019810:Sodium Azide; D011493:Protein Kinase C; D006162:Guanylate Cyclase; D017461:Receptors, Atrial Natriuretic Factor; D006152:Cyclic GMP