Identification and characterization of the phosphorylation sites of the guanylyl cyclase-linked natriuretic peptide receptors A and B.
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.
Potter
L R
LR
Molecular Biology and Virology Laboratory, Salk Institute for Biological Studies, 10010 North Torrey Pines Road, La Jolla, California, 92037, USA. Lincoln@umn.edu
Hunter
T
T
D000409:Alanine / Q000235:genetics / Q000378:metabolism; D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D002460:Cell Line; D018698:Glutamic Acid / Q000235:genetics / Q000378:metabolism; D006162:Guanylate Cyclase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007668:Kidney / Q000166:cytology / Q000378:metabolism; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D010750:Phosphoproteins / Q000737:chemistry; D010766:Phosphorylation; D017461:Receptors, Atrial Natriuretic Factor / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012694:Serine / Q000378:metabolism; D013912:Threonine / Q000378:metabolism; D000099242:Atrial Natriuretic Factor Receptor B
D000409:Alanine; D018698:Glutamic Acid; D006162:Guanylate Cyclase; D010750:Phosphoproteins; D017461:Receptors, Atrial Natriuretic Factor; D011994:Recombinant Proteins; D012694:Serine; D013912:Threonine; D000099242:Atrial Natriuretic Factor Receptor B; C091715:atrial natriuretic factor receptor A