Cloning and functional expression of the bovine natriuretic peptide receptor-B (natriuretic factor R1c subtype.
We describe the isolation of a 3,276 base pair cDNA for the bovine natriuretic peptide receptor-B (NPR-B). Expression of this clone in Cos-P cells demonstrates that it encodes an agonist-dependent guanylyl cyclase. Porcine CNP stimulates the activity of this receptor up to 200-fold with an ED50 of 12 +/- 2 nM, whereas brain natriuretic peptide C-type natriuretic peptide (CNP) and atrial natriuretic factor (ANF) are less efficacious. In addition, ligand binding studies indicate that this receptor exhibits the pharmacology appropriate for the bovine NPR-B. CNP binds to Cos-P cell membranes expressing this clone with a Kd of 13 +/- 1 pM, and natriuretic peptides compete for [125I]-CNP binding with a rank order of pCNP > pBNP > rANF. Thus, the expressed receptor-guanylyl cyclase exhibits the expected pharmacological profile for ligand binding and cyclase activation of the bovine NPR-B receptor.
Department of Pharmacology, University of Montreal, Quebec, Canada.
R
R
Fenrick
K
K
Babinski
N
N
McNicoll
M
M
Therrien
J
J
Drouin
A
A
De Léan
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001921:Brain / Q000378:metabolism*; D002417:Cattle; D002460:Cell Line; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D006152:Cyclic GMP / Q000378:metabolism; D017931:DNA Primers; D018076:DNA, Complementary / Q000302:isolation & purification; D015870:Gene Expression; D015723:Gene Library; D006162:Guanylate Cyclase / Q000096:biosynthesis* / Q000378:metabolism; D006801:Humans; D007668:Kidney; D007700:Kinetics; D008969:Molecular Sequence Data; D020097:Natriuretic Peptide, Brain; D020098:Natriuretic Peptide, C-Type; D009419:Nerve Tissue Proteins / Q000494:pharmacology; D016133:Polymerase Chain Reaction; D011506:Proteins / Q000494:pharmacology; D051381:Rats; D017461:Receptors, Atrial Natriuretic Factor / Q000096:biosynthesis* / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013552:Swine; D014162:Transfection
D017931:DNA Primers; D018076:DNA, Complementary; D009419:Nerve Tissue Proteins; D011506:Proteins; D011994:Recombinant Proteins; D020097:Natriuretic Peptide, Brain; D020098:Natriuretic Peptide, C-Type; D006162:Guanylate Cyclase; D017461:Receptors, Atrial Natriuretic Factor; C073616:atrial natriuretic factor receptor C; D006152:Cyclic GMP