Role of arginine in protein refolding, solubilization, and purification

Biotechnol Prog. 2004 Sep-Oct;20(5):1301-8. doi: 10.1021/bp0498793.

Abstract

Recombinant proteins are often expressed in the form of insoluble inclusion bodies in bacteria. To facilitate refolding of recombinant proteins obtained from inclusion bodies, 0.1 to 1 M arginine is customarily included in solvents used for refolding the proteins by dialysis or dilution. In addition, arginine at higher concentrations, e.g., 0.5-2 M, can be used to extract active, folded proteins from insoluble pellets obtained after lysing Escherichia coli cells. Moreover, arginine increases the yield of proteins secreted to the periplasm, enhances elution of antibodies from Protein-A columns, and stabilizes proteins during storage. All these arginine effects are apparently due to suppression of protein aggregation. Little is known, however, about the mechanism. Various effects of solvent additives on proteins have been attributed to their preferential interaction with the protein, effects on surface tension, or effects on amino acid solubility. The suppression of protein aggregation by arginine cannot be readily explained by either surface tension effects or preferential interactions. In this review we show that interactions between the guanidinium group of arginine and tryptophan side chains may be responsible for suppression of protein aggregation by arginine.

Publication types

  • Research Support, Non-U.S. Gov't
  • Review

MeSH terms

  • Arginine / chemistry*
  • Complex Mixtures / chemistry*
  • Complex Mixtures / isolation & purification*
  • Models, Chemical*
  • Multiprotein Complexes / chemistry*
  • Protein Binding
  • Protein Denaturation
  • Protein Folding
  • Proteins / chemistry*
  • Proteins / isolation & purification*
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / isolation & purification
  • Solubility
  • Structure-Activity Relationship
  • Temperature

Substances

  • Complex Mixtures
  • Multiprotein Complexes
  • Proteins
  • Recombinant Proteins
  • Arginine