Macrophage migration inhibitory factor: isolation from bovine brain.
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.
Cherepkova
O A
OA
Bach Institute of Biochemistry, Russian Academy of Sciences, Moscow, Russia.
Lyutova
E M
EM
Gurvits
B Ya
BY
D000818:Animals; D001923:Brain Chemistry; D002417:Cattle; D002850:Chromatography, Gel / Q000379:methods*; D003600:Cytosol / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D015151:Immunoblotting; D008263:Macrophage Migration-Inhibitory Factors / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D010667:Phenylpyruvic Acids / Q000378:metabolism; D020033:Protein Isoforms
D008263:Macrophage Migration-Inhibitory Factors; D010667:Phenylpyruvic Acids; D020033:Protein Isoforms; C010590:4-hydroxyphenylpyruvic acid