title: Expression, purification, and in vivo activity of atrial natriuretic factor prohormone produced in Escherichia coli.
abstract: Atrial muscles of the heart are known to produce polypeptide hormones called atrial natriuretic factors (ANF) which have potent diuretic and hypotensive action. These hormones are synthesized as a larger protein precursor called pro atrial natriuretic factor or proANF which contains the biologically active ANF sequences at its C-terminus. Rat proANF (representing amino acids -1 to 128 of the coding sequence) was expressed in a soluble form in Escherichia coli. A simple purification procedure was developed which consists of boiling E. coli cell extracts in 1 M acetic acid and subjecting the supernatant to reversed-phase HPLC. The effect of intravenous administration of the purified recombinant proANF on mean arterial blood pressure was examined. The displacement dose-response curves obtained demonstrated that proANF exhibits similar, albeit less potent, physiological activity than ANF.
authors.affiliation: Corporate Research and Development, Biological Sciences, Monsanto Company, Chesterfield, Missouri 63198.
authors.forename: J K
authors.initials: JK
authors.lastname: Gierse
authors.forename: P O
authors.initials: PO
authors.lastname: Olins
authors.forename: C S
authors.initials: CS
authors.lastname: Devine
authors.forename: J D
authors.initials: JD
authors.lastname: Marlay
authors.forename: M G
authors.initials: MG
authors.lastname: Obukowicz
authors.forename: L H
authors.initials: LH
authors.lastname: Mortensen
authors.forename: E G
authors.initials: EG
authors.lastname: McMahon
authors.forename: E H
authors.initials: EH
authors.lastname: Blaine
authors.forename: R
authors.initials: R
authors.lastname: Seetharam
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D009320:Atrial Natriuretic Factor / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology; D001681:Biological Assay; D001794:Blood Pressure / Q000187:drug effects; D002851:Chromatography, High Pressure Liquid; D003001:Cloning, Molecular; D004355:Drug Stability; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D008297:Male; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009154:Mutation; D009318:Natriuresis; D010957:Plasmids; D014176:Protein Biosynthesis; D011498:Protein Precursors / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology; D051381:Rats; D011919:Rats, Inbred Strains; D014158:Transcription, Genetic
chemical_list: D011498:Protein Precursors; D009320:Atrial Natriuretic Factor