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Abstract
A G protein-coupled natriuretic peptide-guanylyl cyclase receptor-B (NPR-B) located in plasma membranes from bovine tracheal smooth muscle shows complex kinetics and regulation. NPR-B was activated by natriuretic peptides (CNP-53 > ANP-28) at the ligand extracellular domain, stimulated by Gq-protein activators, such as mastoparan, and inhibited by Gi-sensitive chloride, interacting at the juxtamembrane domain. The kinase homology domain was evaluated by the ATP inhibition of Mn2+-activated NPR-B, which was partially reversed by mastoparan. The catalytic domain was studied by kinetics of Mn2+/Mg2+ and GTP, and the catalytic effect with GTP analogues with modifications of the 
/γ phosphates and ribose moieties. Most NPR-B biochemical properties remained after detergent solubilization but the mastoparan activation and chloride inhibition of NPR-B disappeared. Our results indicate that NPR-B is a highly regulated nano-machinery with domains acting at cross-talk points with other signal transducing cascades initiated by G protein-coupled receptors and affected by intracellular ligands such as chloride, Mn2+, Mg2+, ATP, and GTP.
Key Words: :
| ABBREVIATIONS: | ||
| ANP-28 | = | atrial NP 1-28 |
| ARM | = | ATP-regulatory module |
| BTSM | = | bovine tracheal smooth muscle |
| CD | = | catalytic domain |
| CNP-22 | = | C-type NP 1-22 |
| CNP-53 | = | C-type NP 1-53 |
| GC | = | guanylyl cyclase |
| GPCR | = | G protein-coupled receptor |
| GPRM | = | G protein regulatory module |
| IMBX | = | isomethylbutylxantine |
| JMD | = | juxtamembrane domain |
| KHD | = | kinase homology domain |
| LExD | = | ligand extracellular domain |
| Mant-GTP | = | N-methylanthraniloyl 2′ GTP |
| NP | = | natriuretic peptide |
| NPR-A or GC-A | = | ANP-sensitive guanylyl cyclase |
| NPR-B or GC-B | = | CNP-sensitive guanylyl cyclase |
| NPR-GC | = | natriuretic peptide receptor guanylyl cyclase |
| NPR-C | = | natriuretic peptide receptor clearance |
| OG | = | octylglucoside |
| OG-GC | = | octylglucoside solubilized guanylyl cyclase |
| PMSF | = | phenylmethylsulfonyl fluoride |
| PTX | = | pertussis toxin |
| sGC | = | soluble guanylyl cyclase |
| SNP | = | sodium nitroprusside |
| TMD | = | transmembrane domain |
| ABBREVIATIONS: | ||
| ANP-28 | = | atrial NP 1-28 |
| ARM | = | ATP-regulatory module |
| BTSM | = | bovine tracheal smooth muscle |
| CD | = | catalytic domain |
| CNP-22 | = | C-type NP 1-22 |
| CNP-53 | = | C-type NP 1-53 |
| GC | = | guanylyl cyclase |
| GPCR | = | G protein-coupled receptor |
| GPRM | = | G protein regulatory module |
| IMBX | = | isomethylbutylxantine |
| JMD | = | juxtamembrane domain |
| KHD | = | kinase homology domain |
| LExD | = | ligand extracellular domain |
| Mant-GTP | = | N-methylanthraniloyl 2′ GTP |
| NP | = | natriuretic peptide |
| NPR-A or GC-A | = | ANP-sensitive guanylyl cyclase |
| NPR-B or GC-B | = | CNP-sensitive guanylyl cyclase |
| NPR-GC | = | natriuretic peptide receptor guanylyl cyclase |
| NPR-C | = | natriuretic peptide receptor clearance |
| OG | = | octylglucoside |
| OG-GC | = | octylglucoside solubilized guanylyl cyclase |
| PMSF | = | phenylmethylsulfonyl fluoride |
| PTX | = | pertussis toxin |
| sGC | = | soluble guanylyl cyclase |
| SNP | = | sodium nitroprusside |
| TMD | = | transmembrane domain |