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Research Article

Characterization of a G Protein -Coupled Guanylyl Cyclase -B Receptor from Bovine Tracheal Smooth Muscle

, , , , , & show all
Pages 269-297 | Published online: 10 Oct 2008
 

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.

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

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