Mutations in B-type natriuretic peptide mediating receptor-A selectivity.
Libraries of monovalent display-phage expressing mutant human B-type natriuretic peptide (hBNP) were used to identify variants that preferentially bind natriuretic peptide receptor-A (NPR-A) compared to receptor-C (NPR-C). Position 19 was a significant determinant of receptor specificity for hBNP display phage. The synthetic hBNP variant S19R had a 265-fold improved NPR-A binding over NPR-C, analogous to the atrial natriuretic peptide (ANP) specificity mutation G16R. Mutation of the last three residues of the hBNP disulfide ring, G23F/L24W/G25R, resulted in about 9-fold improved selectivity. The analogous mutations in ANP decreased NPR-A binding, suggesting divergence in the mechanism of NPR-A recognition.
Schoenfeld
J R
JR
Department of Cardiovascular Research, Genentech, Inc., South San Francisco, CA 94080, USA.
Tom
J Y
JY
Lowe
D G
DG
D000595:Amino Acid Sequence; D009320:Atrial Natriuretic Factor / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology*; D001483:Base Sequence; D001667:Binding, Competitive; D002460:Cell Line; D006152:Cyclic GMP / Q000378:metabolism; D017931:DNA Primers; D015723:Gene Library; D014644:Genetic Variation; D006162:Guanylate Cyclase / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D020097:Natriuretic Peptide, Brain; D016133:Polymerase Chain Reaction; D017461:Receptors, Atrial Natriuretic Factor / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D016415:Sequence Alignment; D013379:Substrate Specificity; D014162:Transfection
D017931:DNA Primers; D011994:Recombinant Proteins; D020097:Natriuretic Peptide, Brain; D009320:Atrial Natriuretic Factor; D006162:Guanylate Cyclase; D017461:Receptors, Atrial Natriuretic Factor; C091715:atrial natriuretic factor receptor A; C073616:atrial natriuretic factor receptor C; D006152:Cyclic GMP