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.
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.
Sharma
R K
RK
Section of Regulatory Biology, Cleveland Clinic Research Institute, OH 44195-5068.
Marala
R B
RB
Duda
T M
TM
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
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