Molecular and Cellular Biochemistry

, Volume 120, Issue 1, pp 69–80 | Cite as

Interaction of atrial natriuretic factor and endothelin-1 signals through receptor guanylate cyclase in pulmonary artery endothelial cells

  • Ravi B. Marala
  • Teresa Duda
  • Rameshwar K. Sharma


The endothelial cell has a unique intrinsic feature: it produces a most potent vasopressor peptide hormone, endothelin (ET-1), yet it also contains a signaling system of an equally potent hypotensive hormone, atrial natriuretic factor (ANF). This raises two related curious questions: does the endothelial cell also contain an ET-1 signaling system? If yes, how do the two systems interact with each other? The present investigation was undertaken to determine such a possibility. Bovine pulmonary artery endothelial (BPAE) cells were chosen as a model system. Identity of the ANF receptor guanylate cyclase was probed with a specific polyclonal antibody to the 180 kDa membrane guanylate cyclase (mGC) ANF receptor. A Western-blot analysis of GTP-affinity-purified endothelial cell membrane proteins recognized a 180 kDa band; the same antibody inhibited the ANF-stimulated guanylate cyclase activity; the ANF-dependent rise of cyclic GMP in the intact cells was dose-dependent. By affinity cross-linking technique, a predominant 55 kDa membrane protein band was specifically labeled with [125I]ET-1. ET-1 treatment of the cells showed a migration of the protein kinase C (PKC) activity from cytosol to the plasma membrane; ET-1 inhibited the ANF-dependent production of cyclic GMP in a dose-dependent fashion with an EC50 of 100 nM. This inhibitory effect was duplicated by phorbol 12-myristate 13-acetate (PMA), a known PKC-activator. The EC50 of PMA was 5 nM. A PKC inhibitor, 1-(5-isoquinolinyl-sulfonyl)-2-methyl piperazine (H-7), blocked the PMA-dependent attenuation of ANF-dependent cyclic GMP formation. These results demonstrate that the 180 kDa mGC-coupled ANF and ET-1 signaling systems coexist in endothelial cells and that the ET-1 signal negates the ANF-dependent guanylate cyclase activity and cyclic GMP formation. Furthermore, these results support the paracrine and/or autocrine role of ET-1.

Key words

atrial natriuretic factor endothelin atrial natriuretic factor receptor guanylate cyclase endothelin receptor protein kinase C 



Atrial Natriuretic Factor




Guanylate Cyclase


membrane Guanylate Cyclase

BPAE cells

Bovine Pulmonary Artery Endothelial cells




3-[cholaimidopropyl)dimethyl-ammonio]-1-Propane Sulfonate


1-(5-isoquinolinyl-sulfonyl)-2-methyl piperazine


Phorbol 12-Myristate


Disuccinimidyl Suberate


Sodium Dodecyl Sulfate


Polyacrylamide Gel Electrophoresis


Hanks Balanced Salt Solution


Bovine Serum Albumin


Phenylmethylsulfonyl Fluoride




[ethylenebis (oxyethylenenitrilo)]tetraacetic Acid


Protein Kinase C


Guanosine Triphosphate


4-(2-hydroxyethyl)-1-Piperazineethanesulfonic acid


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Copyright information

© Kluwer Academic Publishers 1993

Authors and Affiliations

  • Ravi B. Marala
    • 1
  • Teresa Duda
    • 1
  • Rameshwar K. Sharma
    • 1
  1. 1.The unit of regulatory and molecular biologyPennsylvania college of optometryPhiladelphiaUSA

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