Stability of human interferon-beta 1: oligomeric human interferon-beta 1 is inactive but is reactivated by monomerization

Biochim Biophys Acta. 1989 Oct 5;998(2):167-72. doi: 10.1016/0167-4838(89)90269-0.

Abstract

Human interferon-beta 1 is extremely stable is a low ionic strength solution of pH 2 such as 10 mM HCl at 37 degrees C. However, the presence of 0.15 M NaCl led to a remarkable loss of antiviral activity. The molecular-sieve high-performance liquid chromatography revealed that, whereas completely active human interferon-beta 1 eluted as a 25 kDa species (monomeric form), the inactivated preparation eluted primarily as a 90 kDa species (oligomeric form). The specific activity (units per mg protein) of the oligomeric form was approx. 10% of that of the monomeric form. This observation shows that oligomeric human interferon-beta 1 is apparently in an inactive form. When the oligomeric eluate was resolved by polyacrylamide gel containing sodium dodecyl sulphate (SDS), it appeared to be monomeric under non-reducing conditions. Monomerization of the oligomeric human interferon-beta 1 by treatment with 1% SDS, fully regenerated its antiviral activity. These results suggest that the inactivation of the human interferon-beta 1 preparation was caused by its oligomerization via hydrophobic interactions without the formation of intermolecular disulphide bonds. These oligomers can be dissociated by SDS to restore biological activity.

MeSH terms

  • Chromatography, High Pressure Liquid
  • Circular Dichroism
  • Detergents
  • Hot Temperature
  • Humans
  • Hydrogen-Ion Concentration
  • In Vitro Techniques
  • Interferon Type I / ultrastructure*
  • Polymers
  • Protein Conformation
  • Protein Denaturation
  • Structure-Activity Relationship

Substances

  • Detergents
  • Interferon Type I
  • Polymers