Production of an atrial natriuretic peptide variant that is specific for type A receptor.
Receptor-specific variants of atrial natriuretic peptide (ANP) were selected from libraries of filamentous phage particles that displayed single copies of random ANP mutants fused to gene III protein. These ANP variants were differentially selected by binding to immobilized natriuretic peptide receptor A (NPR-A) over competing receptor C (NPR-C) in solution. This method also selected ANP variants with improved secretion expression in Escherichia coli. Several of the identified mutations were combined to produce an efficiently expressed ANP analog that was as potent as wild-type ANP in stimulating NPR-A guanylyl cyclase activity but resistant to inactivation mediated by NPR-C. Such NPR-A-selective analogs should be useful for correlating the various activities of ANP to the relevant receptor and may also be more potent therapeutics in the targeting of NPR-A.
Cunningham
B C
BC
Department of Protein Engineering, Genentech, Inc., South San Francisco, CA 94080.
Lowe
D G
DG
Li
B
B
Bennett
B D
BD
Wells
J A
JA
D000595:Amino Acid Sequence; D009320:Atrial Natriuretic Factor / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D001435:Bacteriophages / Q000235:genetics; D001483:Base Sequence; D004252:DNA Mutational Analysis; D015723:Gene Library; D005821:Genetic Techniques; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009154:Mutation / Q000235:genetics*; D017461:Receptors, Atrial Natriuretic Factor / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D012680:Sensitivity and Specificity
D011993:Recombinant Fusion Proteins; D009320:Atrial Natriuretic Factor; D017461:Receptors, Atrial Natriuretic Factor