Synthesis of recombinant atrial natriuretic peptide (rANP) using hybrid fusion protein-phage fr coat/ANP (CP/ANP).
Recombinant atrial natriuretic peptide (rANP) was expressed in and isolated from E. coli. rANP was purified using HPLC. Amino acid analysis, partial sequencing, and molecular mass were determined. Fused protein was used to rise polyclonal antibodies and to develop of immunoenzymatic assays of rANP and CP/ANP. Experiments were designed to study rANP effects on isolated rabbit aortic strips and to examine hypotensive, diuretic, and natriuretic activity, as well as renal creatinine clearance, in an in vivo rat model. Identity of recombinant and commercial ANP has been confirmed. Physiological activity of CP/ANP has allowed the investigators to predict the conformation of CP/ANP, pro-ANP processing, and the method by which fusion protein interacts with ANP receptors.
Baumanis
V
V
University of Latvia, Biomedical Research and Study Centre, Riga, Latvia.
Jansone
I
I
Skangals
A
A
Mandrika
I
I
Berzins
V
V
D000818:Animals; D000959:Antihypertensive Agents / Q000494:pharmacology; D001011:Aorta / Q000187:drug effects; D009320:Atrial Natriuretic Factor / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology; D002213:Capsid / Q000235:genetics* / Q000302:isolation & purification; D036022:Capsid Proteins; D002851:Chromatography, High Pressure Liquid; D004231:Diuresis / Q000187:drug effects; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000378:metabolism; D005919:Glomerular Filtration Rate / Q000187:drug effects; D066298:In Vitro Techniques; D008666:Metalloendopeptidases / Q000378:metabolism; D009126:Muscle Relaxation / Q000187:drug effects; D009131:Muscle, Smooth, Vascular / Q000187:drug effects; D011817:Rabbits; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology
D000959:Antihypertensive Agents; D036022:Capsid Proteins; D011993:Recombinant Fusion Proteins; C085782:coat protein, Enterobacteria phage fr; D009320:Atrial Natriuretic Factor; D008666:Metalloendopeptidases; C039830:peptidyl-Lys metalloendopeptidase