Ambrosio A L, Iglesias M M, Wolfenstein-Todel C
Instituto de Química y Fisicoquímica Biológicas, (UBA-CONICET), Facultad de Farmacia y Bioquímica, Buenos Aires, Argentina.
Glycoconj J. 1997 Nov;14(7):831-6. doi: 10.1023/a:1018538004923.
The heparin-binding lectin complex from ovine placental cotyledons was purified by affinity chromatography on heparin-agarose column. It showed three protein bands, which had molecular weights of 13000, 15000 and 17000 by sodium dodecylsulfate-polyacrylamide gel electrophoresis, and the presence of DNA by agarose gel electrophoresis. The protein components of the complex were separated by reverse-phase HPLC. The minimum inhibitory concentrations of glycosaminoglycans were significantly different for the lectin complex and the separated proteins, suggesting affinity changes upon DNA binding. The haemagglutinating activity specificity allowed the characterization of the fraction with a molecular weight of 13000 as the heparin-binding lectin. This protein was identified as histone H4 by internal sequencing, thus showing that this is the histone responsible for the heparin-binding property of the complex. The accompanying proteins were tentatively identified as histones H2A and H2B.
通过在肝素 - 琼脂糖柱上进行亲和层析，从绵羊胎盘子叶中纯化出肝素结合凝集素复合物。经十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳显示，它呈现出三条蛋白带，分子量分别为13000、15000和17000，且经琼脂糖凝胶电泳表明存在DNA。该复合物的蛋白质组分通过反相高效液相色谱进行分离。凝集素复合物和分离出的蛋白质对糖胺聚糖的最小抑制浓度存在显著差异，这表明DNA结合后亲和力发生了变化。血细胞凝集活性特异性使得能够将分子量为13000的组分鉴定为肝素结合凝集素。通过内部测序，该蛋白质被鉴定为组蛋白H4，因此表明这就是负责复合物肝素结合特性的组蛋白。伴随的蛋白质初步鉴定为组蛋白H2A和H2B。
