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Macrophage Migration Inhibitory Factor: Isolation from Bovine Brain

O. A. Cherepkova, E. M. Lyutova, and B. Ya. Gurvits*

Bach Institute of Biochemistry, Russian Academy of Sciences, Leninsky pr. 33, 119071 Moscow, Russia; fax: (7-495) 954-2732; E-mail: bella@inbi.ras.ru

* To whom correspondence should be addressed.

Received September 1, 2004; Revision received December 22, 2004
The purification of macrophage migration inhibitory factor (MIF) from bovine brain cytosol and its partial characterization are reported. A rapid and relatively simple method for MIF isolation was developed based mainly on size-exclusion chromatography on Toyopearl TSK polymer having a tendency to adsorb MIF as compared to elution of other proteins with similar molecular weights. The method gives a high yield of MIF (0.1 mg homogenous protein per g wet tissue). The retardation is conveniently utilized to achieve good separations of MIF from other proteins of similar molecular weights. The isolated protein was identified as MIF by SDS-electrophoresis, immunoblotting, sequencing of the N-terminal amino acid residues, and also by determination of keto-enol tautomerase activity that is characteristic of MIF with p-hydroxyphenylpyruvic acid as a substrate.
KEY WORDS: macrophage migration inhibitory factor, gel filtration, keto-enol tautomerase

DOI: 10.1134/S0006297906010111