Cherepkova O A, Lyutova E M, Gurvits B Ya
Bach Institute of Biochemistry, Russian Academy of Sciences, Moscow, Russia.
Biochemistry (Mosc). 2006 Jan;71(1):73-8. doi: 10.1134/s0006297906010111.
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.
本文报道了从牛脑细胞质中纯化巨噬细胞移动抑制因子（MIF）及其部分特性。主要基于在Toyopearl TSK聚合物上的尺寸排阻色谱法，开发了一种快速且相对简单的MIF分离方法，与洗脱其他分子量相似的蛋白质相比，该聚合物倾向于吸附MIF。该方法可获得高产率的MIF（每克湿组织0.1毫克纯蛋白）。这种阻滞作用便于将MIF与其他分子量相似的蛋白质进行良好分离。通过SDS电泳、免疫印迹、N端氨基酸残基测序以及以对羟基苯丙酮酸为底物测定MIF特有的酮-烯醇互变异构酶活性，鉴定出分离的蛋白质为MIF。
