Expression, purification, and in vivo activity of atrial natriuretic factor prohormone produced in Escherichia coli

Arch Biochem Biophys. 1989 Jun;271(2):441-6. doi: 10.1016/0003-9861(89)90294-4.

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.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Atrial Natriuretic Factor* / genetics
  • Atrial Natriuretic Factor* / isolation & purification
  • Atrial Natriuretic Factor* / pharmacology
  • Biological Assay
  • Blood Pressure / drug effects
  • Chromatography, High Pressure Liquid
  • Cloning, Molecular
  • Drug Stability
  • Escherichia coli / genetics
  • Escherichia coli / metabolism*
  • Hydrogen-Ion Concentration
  • Male
  • Molecular Sequence Data
  • Molecular Weight
  • Mutation
  • Natriuresis
  • Plasmids
  • Protein Biosynthesis
  • Protein Precursors* / genetics
  • Protein Precursors* / isolation & purification
  • Protein Precursors* / pharmacology
  • Rats
  • Rats, Inbred Strains
  • Transcription, Genetic

Substances

  • Protein Precursors
  • Atrial Natriuretic Factor