Abstract
The binding of atrial natriuretic peptide and C-type natriuretic peptide to the guanylyl cyclase-linked natriuretic peptide receptors A and B (NPR-A and NPR-B), respectively, results in decreases in extracellular volume, vascular tension and cell proliferation. Both NPR-A and NPR-B are extensively phosphorylated in resting cells and receptor dephosphorylation is correlated with ligand-induced homologous desensitization. To understand the role of phosphorylation in the regulation of these receptors, we identified the in vivo phosphorylation sites of NPR-A and NPR-B and found that the phosphorylation of multiple sites within their kinase homology domains is absolutely required for their activation. In this review, we give a detailed description of the phosphopeptide mapping techniques that were used to identify and characterize these sites and discuss the potential pitfalls that are associated with them.
Copyright 1999 Academic Press.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Alanine / genetics
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Alanine / metabolism
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Amino Acid Motifs
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Amino Acid Sequence
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Amino Acid Substitution
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Cell Line
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Glutamic Acid / genetics
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Glutamic Acid / metabolism
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Guanylate Cyclase / chemistry
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Guanylate Cyclase / genetics
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Guanylate Cyclase / metabolism*
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Humans
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Kidney / cytology
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Kidney / metabolism
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Molecular Sequence Data
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Mutagenesis, Site-Directed
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Mutation
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Phosphoproteins / chemistry
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Phosphorylation
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Receptors, Atrial Natriuretic Factor / chemistry
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Receptors, Atrial Natriuretic Factor / genetics
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Receptors, Atrial Natriuretic Factor / metabolism*
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Recombinant Proteins / genetics
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Recombinant Proteins / metabolism
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Serine / metabolism
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Threonine / metabolism
Substances
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Phosphoproteins
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Recombinant Proteins
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Threonine
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Glutamic Acid
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Serine
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Guanylate Cyclase
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Receptors, Atrial Natriuretic Factor
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atrial natriuretic factor receptor A
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atrial natriuretic factor receptor B
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Alanine