Calcium-dependent dephosphorylation mediates the hyperosmotic and lysophosphatidic acid-dependent inhibition of natriuretic peptide receptor-B/guanylyl cyclase-B

Regine Potthast, Sarah E. Abbey-Hosch, Laura K. Antos, Jonathan S. Marchant, Michaela Kuhn, Lincoln R. Potter

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

C-type natriuretic peptide binding to natriuretic peptide receptor-B (NPR-B) stimulates cGMP synthesis, which regulates vasorelaxation, cell proliferation, and bone growth. Here, we investigated the mechanistic basis for hyperosmotic and lysophosphatidic acid-dependent inhibition of NPR-B. Whole cell cGMP measurements and guanylyl cyclase assays indicated that acute hyperosmolarity decreased NPR-B activity in a reversible, concentration- and time-dependent manner, whereas chronic exposure had no effect. Acute hyperosmolarity elevated intracellular calcium in a concentration-dependent fashion that paralleled NPR-B desensitization. A calcium chelator, but not a protein kinase C inhibitor, blocked both calcium elevations and desensitization. Hyperosmotic medium stimulated NPR-B dephosphorylation, and the receptor was rapidly rephosphorylated and resensitized when the hypertonic media was removed. Lysophosphatidic acid also inhibited NPR-B in a calcium- and phosphorylation-dependent process, consistent with calcium being a universal regulator of NPR-B. The absolute requirement of dephosphorylation in this process was demonstrated by showing that a receptor with glutamates substituted at all known NPR-B phosphorylation sites is unresponsive to hyperosmotic stimuli. This is the first study to measure the phosphorylation state of an endogenous guanylyl cyclase and to link intracellular calcium elevations with its dephosphorylation.

Original languageEnglish (US)
Pages (from-to)48513-48519
Number of pages7
JournalJournal of Biological Chemistry
Volume279
Issue number47
DOIs
StatePublished - Nov 19 2004

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Guanylate Cyclase
Calcium
Phosphorylation
C-Type Natriuretic Peptide
Glutamates
Protein C Inhibitor
Bone Development
Glutamate Receptors
Cell proliferation
Protein Kinase Inhibitors
Vasodilation
Protein Kinase C
lysophosphatidic acid
atrial natriuretic factor receptor B
Assays
Bone
Cell Proliferation

Cite this

Calcium-dependent dephosphorylation mediates the hyperosmotic and lysophosphatidic acid-dependent inhibition of natriuretic peptide receptor-B/guanylyl cyclase-B. / Potthast, Regine; Abbey-Hosch, Sarah E.; Antos, Laura K.; Marchant, Jonathan S.; Kuhn, Michaela; Potter, Lincoln R.

In: Journal of Biological Chemistry, Vol. 279, No. 47, 19.11.2004, p. 48513-48519.

Research output: Contribution to journalArticle

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