The structure and physicochemical properties of rat liver macrophage migration inhibitory factor

Biochim Biophys Acta. 1995 Feb 22;1247(1):159-62. doi: 10.1016/0167-4838(94)00215-3.


We expressed rat macrophage migration inhibitory factor (rMIF) in E. coli using the cDNA isolated from a rat liver cDNA library. rMIF specifically bound glutathione (dissociation constant = 500 microM). We purified rMIF homogeneously on SDS-PAGE by S-hexylglutathione Sepharose affinity column chromatography and Sephadex G-100 column chromatography. The amino-acid sequence of rMIF was highly homologous to that of human MIF from a T-cell line; only a single amino-acid residue was substituted if conservative amino-acid substitutions were involved. The molecular weight of rMIF was calculated to be 12.4 kDa and 23.6 kDa by SDS-PAGE and analytical ultracentrifugation, respectively. Thus, it was concluded that the native rMIF formed a homodimeric structure. Proton nuclear magnetic resonance (1H-NMR) study revealed that rMIF was less thermostable (the denaturing temperature was from 50-60 degrees C) than human MIF (the denaturing temperature is about 80 degrees C (Nishihira et al. (1993) Biochem. Mol. Biol. Int. 31, 841-850). The secondary structure of rMIF evaluated by 1H-NMR experiments revealed that the contents of alpha-helix, beta-strand, and coil were 13.8%, 55.6%, and 30.6%, respectively.

MeSH terms

  • Animals
  • Escherichia coli / metabolism
  • Liver / metabolism*
  • Macrophage Migration-Inhibitory Factors / biosynthesis
  • Macrophage Migration-Inhibitory Factors / isolation & purification*
  • Macrophages / metabolism*
  • Magnetic Resonance Spectroscopy
  • Molecular Weight
  • Protein Structure, Secondary
  • Rats
  • Ultracentrifugation


  • Macrophage Migration-Inhibitory Factors